Analysis by Confocal Microscopy of the Structure of Cambium in the Hardwood Kalopanax pictus

An understanding of the morphology and the developmental changesin the shapes and dimensions of cambial cells requires three-dimensional(3-D) analysis of thick slices of tissue. We devised a simpleprotocol using confocal laser scanning microscopy (CLSM), withsafranin and acridine orange as fluorescent dyes and glycerolas the clearing and mounting medium, to examine the 3-D structureof the dormant cambium in Kalopanax pictus, a ring-porous hardwood.Optical sections and high contrast images provided clear informationabout the shapes and nuclear status of cambial cells, whichhave previously been difficult to determine using conventionalmicroscopy. The axially-oriented cambial cells were found tovary in shape, in particular around the rays, and were not alwaystypically fusiform. We evaluated the reliability of our methodby comparing results with those of a parallel study of the samematerial by standard analysis of serial sections of epoxy-embeddedspecimens. The images of optical sections obtained by CLSM wereof high quality and similar to images obtained by conventionallight microscopy of semi-thin mechanical sections. Use of theconfocal microscope provided a quick and easy method for visualizationof the structure of the cambium in thick hand-cut sections andfor studies of the developmental changes in cells from the cambiumto the xylem. Copyright 2000 Annals of Botany Company Confocal laser scanning microscopy, Kalopanax pictus, three-dimensional reconstruction, vascular cambium     

Studies on Mature Seeds of Cuscuta pedicellata and C. campestris by Electron Microscopy

The seeds of Cuscuta pedicellata have been investigated by transmissionand scanning electron microscopy. Additional observations havebeen made on seeds of C. campestris by SEM only. The seed coatconsists of an outer single epidermis, two different palisadelayers, and an inner multiparenchyma layer. The outer epidermalwall in C. pedicellata has a thick cuticle and zones rich inpectic substances. The thicker ‘U-shaped’ cell wallsin the outer palisade layer are strengthened by a wall layerof hemicellulose. The inner palisade layer has thick walledcells with a ‘light line’. The inner cell wall ofthe compressed multiparenchyma layer has a thin cuticle. A fairlythick cuticle is positioned directly on the endosperm surface.The aleurone cell walls are different from the remaining endospermwalls. The latter are thick and believed to be of galactomannans.There is a ‘clear’ zone between the plasmalemmaand the cell wall in the aleurone cells. The embryo cells arepacked with lipids and proteins. In Cuscuta campestris mostendosperm has been absorbed during the seed development. Theembryo apex has two minute leaf primordia. The features of theCuscuta seeds are discussed in relation to functional and environmentalconditions. Cuscuta pedicellata, Cuscuta campestris, seed, seed coat, cuticle, cell walls, endosperm, aleurone cells, galactomannan, embryo, TEM, SEM     

Cellular Structure and Light Absorbtion in Leaves of Frithia pulchra (Mesembryanthemaceae)

In order to quantify the structural differences between celltypes of leaves from a ‘ window’ plant, an ultrastructuralmorphometric analysis was made of the epidermal, window andchlorenchyma tissues of Frithia pulchra. Epidermal cells arethe largest cells found in Frithia leaves and are characterizedby the presence of a thick outer tangential cell wall and numerousvacuolar inclusions. Epidermal tissue has an optical densityof 0.30. The transparent window tissue (i.e. optical density= 0.08) has a uniform ultrastructure throughout the length ofthe leaf. The vacuome comprises aproximately 97 per cent ofthe protoplasmic volume of window cells. Chlorenchyma cellspossess thin cell walls and are surrounded by numerous intercellularspaces. Cells of the apical chlorenchyma tissue possess approximately30 plastids per cell. These chloroplasts have an average individualvolume of 220 µm². Cells of the basal chlorenchyma tissuecontain chloroplasts that are five to six times smaller andmore numerous than those in cells of the apical chlorenchyma.The increased volume of chloroplasts in the apical comparedwith basal chlorenchyma cells (i.e. 31.4 and 20.2 per cent ofthe protoplasm, respectively) is positively correlated withtheir optical densities of 1.46 and 0.97, respectively. Frithia pulchra, stereology, leaf, light absorption, window plant     

Leaf Anatomy, Emphasizing Unusual 'Concertina' Mesophyll Cells, of Two East African Legumes (Caesalpinieae, Caesalpinioideae, Leguminosae)

Cordeauxia edulis (Somalia and Ethiopia), andStuhlmannia moavii(Tanzania, Kenya and Madagascar) are evergreen shrubs or smalltrees of dry areas. They have similar leaf anatomy as revealedby resin sectioning and scanning electron microscopy. The cuticleis extremely thick and all vascular bundles lack bundle sheathextensions. The most unusual feature is the mesophyll, threeto seven layers consisting entirely of cylindrical palisadecells with lateral walls capable of changing vertical lengthby folding in a concertina-like manner. The matching outwardfolds of two adjacent cells always remain attached by meansof a row of wall thickenings (‘pegs’). The pegscan elongate, especially so between the widely separated mesophyllcells that occupy the substomatal chamber area. The unattachedflexible inward wall folds enable these ‘concertina’cells to shorten or lengthen vertically without disrupting cellinterconnections in the interior of each relatively long-livedleaf as it periodically loses and gains water. Concertina cellsmay be an anatomical adaptation allowing these leaves to remainevergreen and survive extended periods of drought and yet tostore water quickly when it becomes available. Leguminosae; Caesalpinioideae; Cordeauxia ; Stuhlmannia ; ‘concertina’ mesophyll cells; desert adaptation; hollow glandular trichomes; leaf anatomy; wall thickenings     

Low Temperature Affects Pattern of Leaf Growth and Structure of Cell Walls in Winter Oilseed Rape (Brassica napus L., var. oleifera L.)

Three-week acclimation of winter oilseed rape (Brassica napusL. var. oleifera L.) plants in the cold (2 °C) resultedin a modified pattern of leaf cell enlargement, indicated bythe increased thickness of young leaf blades and modified dimensionsof mesophyll cells, as compared with non-acclimated tissuesgrown at 20/15 °C (day/night). The thickness of leaf cellwalls also increased markedly during cold acclimation but itdecreased in response to a transient freezing event (5 °Cfor 18 h followed by 6 or 24 h at 2 °C, in the dark). Cellwalls of the upper (adaxial) epidermis were most affected. Theirultrastructure was modified by cold and freezing treatmentsin different ways, as revealed by electron microscopy. Possiblereasons for the cold- and freezing-induced modifications inthe leaf and cell wall morphology and their role in plant acclimationto low temperature conditions are discussed. Copyright 1999Annals of Botany Company Acclimation, Brassica napus var. oleifera, cell wall ultrastructure, cold, freezing, leaf structure, winter oilseed rape.     

The Anatomy, Histochemistry and Ultrastructure of Stigmas and Styles in Commelinaceae

The anatomy and ultrastructure of stigmas in 37 species of 13genera of Commelinaceae are described. The stigmas are papillate,papillae forming a dense fringe of cells around the mouth ofthe stylar canal in most species. The papillar cell wall iscovered by an unstructured cuticle of variable thickness andis of variable thickness because of small wall ingrowths. Thecuticle and the external surface of the papillar cell wall arevariably disrupted, particularly in the mid and basal regionsof the cell. This was not found in species of the genus Aploleiaor Callisia. The cell cytoplasm possesses all major organellesexcept chloroplasts and each cell is vacuolate. In all species except Aploleia mulitiflora the style comprisesan epidermis, a cortex and a hollow, tripartite canal whichis continuous into the ovary cavity. The three vascular strandsare positioned at the apex of each canal lobe. The canal cellsare elongate and tabular and the wall abutting the canal hasingrowths. The style in Aploleia is solid and the transmittingtissue comprises cells whose walls are electron opaque. Thecytoplasms of both types of cell are similar in content althoughthere is a single, large vacuole in canal cells and many smallvacuoles in transmitting tissue. The morphology, position and histochemistry of stigmatic andstylar exudate was similar in all ‘wet’ stigmas.Most of the exudate originates from the stylar canal althoughsignificant contributions are made by the papillae in stigmasof Coleotrype, Dichorisandra and Thyrsanthemum. There is no apparent relationship between stigma structure andthe presence of self-incompatibility. Stigma papillae, stylar canal, transmitting tissue, Commelinaceae     

Three-dimensional imaging of tumor angiogenesis

OBJECTIVE: To three-dimensionally visualize the microvessel environment of tumor angiogenesis by confocal laser scanning microscopy (CLSM). STUDY DESIGN: To reveal underlying mechanisms of tumor angiogenesis, a 7, 12-dimethylbenz(a) anthracene-induced rat cancer model was used. For demonstrating tumor vasculature, fluorescence injection method (FITC-conjugated gelatin solution) was employed. FITC gelatin was injected into the left ventricle of the rat heart. After complete perfusion, the mammary glands were resected, fixed under ice cold conditions and subjected to immunohistochemistry (IHC) for tumor cells. The LSM-410 (Carl Zeiss, Jena, Germany) was employed on thick sections (300-2,000 microns) to elucidate detailed microvessel networks (MVN) and tumor cells. RESULTS: Tumor vasculature on thick sections was clearly detected by CLSM at the maximum focus depth of 2,000 microns. Three-dimensional (3-D), reconstructed images of normal mammary glands showed regular and linear MVN. In DMBA-induced mammary cancer, vascular density of MVN was markedly increased and showed an anastomosing, irregular MVN pattern. Furthermore, focal segmentation and tortuous, branching patterns of microvessels were also seen. CONCLUSION: Application of the fluorescence injection method and IHC using CLSM was very useful for studying the 3-D relationship between tumor angiogenesis and neoplastic epithelial changes. These results suggest that application of this technique is ideal for studying 3-D imaging of tumor angiogenesis.     

Autolysis Promotes the Extension Capacity of Zea mays Coleoptile Cell Walls in Response to Acid pH Solutions

The relationship between autolytic degradation of ß(1–3),(1–4)-D-glucanand acid pH-induced extension of isolated Zea mays cell wallshas been investigated using a constant-load extension technique.Acidic buffer (4.5) was able to induce an additional extension(E_a) on cell walls already extended at pH 6.8 buffer under a20 g-mass load, indicating that the additional extension (E_a)was the parameter that better represented the effect of thedifferent treatments on the mechanical properties of maize coleoptilecell walls. The additional extension in response to acidic pHwas higher when cell walls had been previously autolysed for24 h at pH 5.5. Furthermore, the acid-pH effect was dependenton the presence during the constant load extension of some thermo-labilefactors, suggesting the participation of expansins. Acid pHincreased E_a of native cell walls through an increase in theplastic extension (E_p) in agreement with a one step mechanismleading directly to irreversible (plastic) wall extension assuggested by Cosgrove (1977). The autolytic degradation of ß(1–3),(1–4)-D-glucan was also able to modify the mechanicalproperties of maize coleoptile cell walls increasing its elasticextension (E_e) in response to pH 4.5 buffer but that modificationonly leads to an increase in wall extension when expansins areactive, suggesting a cooperation between ß-glucanturnover and expansin action. (Received August 5, 1998; Accepted March 16, 1999)     

Voigt and Reuss Models for Predicting Changes in Young's Modulus of Dehydrating Plant Organs

A general modelling approach was used to predict the changesresulting from dehydration in the Young's modulus (E) of a tereteorgan with a simple anatomy (e.g. hypodermis of thick-walledtissue surrounding a parenchymatous core of ground tissue).Two general anatomical models were investigated: (1) an ‘apoplastmodel’ in which each cross section through the organ wasconsidered as a composite elastic material consisting of a solid(cell wall) and liquid (protoplasm) phase; and (2) a ‘core-rindmodel’ in which the organ consisted of a thick-walledand a thin-walled tissue. For each of these two anatomical models,two composite material models were considered, i.e. a Voigtor Reuss equation was used to predict the changes in E attendingdehydration. The predictions from the variants of the generalmodel were evaluated on the basis of observed changes in E ascylindrical segments of the pseudopetioles of Spathiphyllumwere allowed to desiccate at room temperature. Statistical comparisonsbetween predicted and observed values of E revealed that oneof the simple variants of the model, the ‘Voigt apoplastmodel’, was the most successful in predicting the observedtrend seen in the changes in E. However, when the Voigt andReuss apoplast models were combined, the ‘hybrid’model provided estimates of changes in E that were statisticallyindistinguishable from those observed. Based on the hybrid model,it was estimated that roughly 76.7% of the tissues with a representativeSpathiphyllum pscudopetiole operated according to a Voigt apoplastmodel. Young's modulus, dehydration, plant tissues     

STEREOLOGICAL METHODS FOR MEASURING INTERNAL LEAF STRUCTURE VARIABLES

A recent increase in research into ecological leaf anatomy is based partly on the effects of leaf structure upon photosynthesis. Hence, it is desirable to have efficient methods for measuring various aspects of internal leaf anatomy, such as percent air space and cell wall area per unit volume of tissue. Stereological methods make use of point counts and of counts of the number of intersections of sampling lines with tissue outlines on planar sections. Their practical use for measuring plant tissues is described, with special reference to highly directional tissue like palisade mesophyll and wood. In particular, the method for estimating cell wall area per unit volume requires the choice of a numerical factor which varies with tissue type. The factor has values of 2 for unoriented tissues, pI/2 for cylinders cut in cross section, and (p/2)² for cylinders. in longitudinal section. An interpolating function to derive a value of the factor for tissues of intermediate type is also described. Leaves from two species of composites from contrasting habitats are compared, to demonstrate the usefulness of the methods.     

Confocal DNA cytometry: a contour-based segmentation algorithm for automated three-dimensional image segmentation

BACKGROUND: Confocal laser scanning microscopy (CLSM) presents the opportunity to perform three-dimensional (3D) DNA content measurements on intact cells in thick histological sections. So far, these measurements have been performed manually, which is quite time-consuming. METHODS: In this study, an intuitive contour-based segmentation algorithm for automatic 3D CLSM image cytometry of nuclei in thick histological sections is presented. To evaluate the segmentation algorithm, we measured the DNA content and volume of human liver and breast cancer nuclei in 3D CLSM images. RESULTS: A high percentage of nuclei could be segmented fully automatically (e.g., human liver, 92%). Comparison with (time-consuming) interactive measurements on the same CLSM images showed that the results were well correlated (liver, r = 1.00; breast, r = 0.92). CONCLUSIONS: Automatic 3D CLSM image cytometry enables measurement of volume and DNA content of large numbers of nuclei in thick histological sections within an acceptable time. This makes large-scale studies feasible, whereby the advantages of CLSM can be exploited fully. The intuitive modular segmentation algorithm presented in this study detects and separates overlapping objects, also in two-dimensional (2D) space. Therefore, this algorithm may also be suitable for other applications.     

Ultrastructural Changes in Coconut Calli Associated with the Acquisition of Embryogenic Competence

Ultrastructural studies of 2,4-D (2,4 dichlorophenoxyaceticacid) induced coconut calli and of untreated controls enabledus to characterize early events in cellular reorganization leadingto embryogenic cell individualization and subsequent developmentinto proembryos. Embryogenic cells were characterized by specialfeatures that chiefly affected the nucleus, cytoplasm and cellwall: deep invaginations of the nuclear envelope, proliferationof dictyosomes, with emission of Golgi vesicles, directly relatedto an increase in cell wall thickness. Modification of the cellwall structure was studied and particular attention was paidto the cytolocalization of ß-1,4-glucans, and of calloseand pectin epitopes, using gold-conjugated probes. The firstchanges (detected 7–14 d after 2,4-D increase) involvedthe closure of plasmodesmata, breaking of symplastic continuity,and callose deposition. The acquisition of embryogenic competencewas linked to the appearance of an outer layer of fibrillarmaterial containing pectin epitope (mainly un-methyl-esterified),fully coating the embryogenic cells (21 d after the inductiontreatment). Some of the ultrastructural changes observed duringthe reprogramming of somatic cells towards embryogenesis canbe likened to those accompanying the maturation of female gametecells in many plant species. The possible significance of theseobservations is discussed. Copyright 2001 Annals of Botany Company Callose, cell wall structure, Cocos nucifera L., cytological events, embryogenic cells, pectin, somatic embryogenesis     

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     

Glandular Trichomes in Satureja thymbra Leaves

The leaves of the aromatic plant Satureja thymbra have numerousglandular trichomes of two morphologically distinct types glandularhairs and glandular scales Investigations of the anatomy ofthese glandular trichomes with serial thick sections revealedthat the glandular hairs consist of three cells a foot, stalkand head cell Glandular scales also have a unicellular footand stalk Their heads, however, are composed of 12 cells Fourof these cells are small, occupying the central region of thehead, whereas the remainder are large and peripherally arrangedMorphometric analysis showed that, in leaf surface view, glandularscales are about 17-fold larger than glandular hairs In addition,glandular scales were found to occupy 5 7 % of the entire leafsurface area In each glandular scale the total amount of essentialoil, contained within both the subcuticular space and the interiorof the secretory cells, was calculated to be 2 51 x 10^–4mm³ The volume of the essential oil produced by all glandularscales on a single mature leaf was correspondingly determinedto be 0.059 mm³ Finally, the theoretical essential oil yieldof 100 g dry leaves of S thymbra was estimated to be 3 54 %(secretory activity of glandular scales only) Satureja thymbra, glandular trichomes, morphology, morphometry     

Anatomical considerations related to photosynthesis in cotton (Gossypium hirsutum L.) leaves, bracts, and the capsule wall

Light and electron microscopy was used to relate histologicaland ultrastructural differences of cotton (Gossypium hirsutumL.) leaves, bracts, and capsule walls to their different photosyntheticactivities. Light microscopy revealed that the leaf thicknesswas approximately 152µm, had a well-defined internal organizationwith elongated palisade mesophyll cells and loosely packed spongymesophyll cells. In contrast, the bract was thinner (111 µm),lacked a defined palisade layer, and was largely composed ofinternal air spaces. The capsule wall was very thick (1013µm)and composed of numerous tightly packed, paren-chymatous corticalcells with little or no intercellular air space. Chloroplastswith well-defined granal stacks and extensive stroma lamellaewere observed in each of these three tissues, however, theirdensity was always greater in the palisade cells of the leafcompared to spongy mesophyll cells of the bract and the parenchymatouscells of the capsule wall. The low rates of photosynthesis inthe bracts and the capsule wall were associated with the internalorganization of these tissues. Key words: Cotton, photosynthesis, anatomy, cuticle, tissues     

ZmXTH1, a new xyloglucan endotransglucosylase/hydrolase in maize, affects cell wall structure and composition in Arabidopsis thaliana

Xyloglucan endotransglucosylase/hydrolases (XTHs; EC 2.4.1.207and/or EC 3.2.1.15 [EC] 1) are enzymes involved in the modificationof cell wall structure by cleaving and, often, also re-joiningxyloglucan molecules in primary plant cell walls. Using a poolof antibodies raised against an enriched cell wall protein fraction,a new XTH cDNA in maize, ZmXTH1, has been isolated from a cDNAexpression library obtained from the elongation zone of themaize root. The predicted protein has a putative N-terminalsignal peptide and possesses the typical domains of this enzymefamily, such as a catalytic domain that is homologous to thatof Bacillus macerans β-glucanase, a putative N-glycosylationmotif, and four cysteine residues in the central and C terminalregions of the ZmXTH1 protein. Phylogenetic analysis of ZmXTH1reveals that it belongs to subgroup 4, so far only reportedfrom Poaceae monocot species. ZmXTH1 has been expressed in Pichiapastoris (a methylotrophic yeast) and the recombinant enzymeshowed xyloglucan endotransglucosylase but not xyloglucan endohydrolaseactivity, representing the first enzyme belonging to subgroup4 characterized in maize so far. Expression data indicate thatZmXTH1 is expressed in elongating tissues, modulated by cultureconditions, and induced by gibberellins. Transient expressionassays in onion cells reveal that ZmXTH1 is directed to thecell wall, although weakly bound. Finally, Arabidopsis thalianaplants expressing ZmXTH1 show slightly increased xyloglucanendohydrolase activity and alterations in the cell wall structureand composition. Key words: Cell elongation, cell wall, plant transformation, XEH, XET, XTH, Zea mays     

Three-dimensional electron microscopy of ribosomal chromatin in two higher plants: a cytochemical, immunocytochemical, and in situ hybridization approach

We report the 3-D arrangement of DNA within the nucleolar subcomponents from two evolutionary distant higher plants, Zea mays and Sinapis alba. These species are particularly convenient to study the spatial organization of plant intranucleolar DNA, since their nucleoli have been previously reconstructed in 3-D from serial ultra-thin sections. We used the osmium ammine-B complex (a specific DNA stain) on thick sections of Lowicryl-embedded root fragments. Immunocytochemical techniques using anti-DNA antibodies and rDNA/rDNA in situ hybridization were also applied on ultra-thin sections. We showed on tilted images that the OA-B stains DNA throughout the whole thickness of the section. In addition, very low quantities of cytoplasmic DNA were stained by this complex, which is now the best DNA stain used in electron microscopy. Within the nucleoli the DNA was localized in the fibrillar centers, where large clumps of dense chromatin were also visible. In the two plant species intranucleolar chromatin forms a complex network with strands partially linked to chromosomal nucleolar-organizing regions identified by in situ hybridization. This study describes for the first time the spatial arrangement of the intranucleolar chromatin in nucleoli of higher plants using high-resolution techniques.     

Observations on the Fine Structure of Plant Cell Walls II. The Microfibrillar Framework of the Parenchymatous Cell Wall in Cucurbita

The microfibrillar framework of parenchymatous walls in Cucurbitawas observed in petioles treated so as to remove various non-cellulosiccell wall components. Such extraction typically results in separationof the microfibrillar components into concentric lamellae. Thenumber and thickness of these lamellae vary according to theage and type of cell wall. The microfibrils appear to be orientatedwithin the plane of their lamellae but the orientation may varyin successive lamellae, and in many walls the crossed polylamellatecondition was detected. The collenchyma—and the outerepidermal cell walls show an alternation of lamellae with almostvertical microfibrils with those with a practically transverseorientation. In ordinary parenchymatous walls the alternationis not so extreme and is revealed only by the occasional presenceof the ‘herring bone pattern’ in non-radial sections.As a rule the lamellae are continuous around the circumferenceof a cell though individual lamellae may vary in thickness andsometimes appear to ‘fade out’. The present observationssuggest that growth of the primary wall occurs by depositionof microfibrils in successive lamellae thus confirming the basicpremise of the multinet theory of growth.     

Studies on the Growth in Culture of Plant Cells: XXI FINE STRUCTURAL FEATURES OF ACER PSEUDOPLATANULS L. CELLS RESPONDING TO 2,4-DICHLOROPHENOXYACETIC ACID WITHLDRAWAL IN TURBIDOSTAT CULTURE

A culture of Acer pseudoplatanus L. grown in the presence ofan equilibrium level of 2,4-dichlorophenoxyacetic acid (2,4-D)of 1?5 ? 10^–7 M (state IV culture) showed, in comparisonwith one of a similar specific growth rate but in which theequilibrium level of 2,4-D was 2?3 ? 10^–6 M (state Iculture), an enhanced degree of cell aggregation, enhanced meancell volume, and the presence of cells giving a generalizedlignin reaction with extracellular lignin-positive material.The state IV culture showed a proportion (10–15 per cent)of cells having ultrastructural features not observed in thestate I culture. Some of the cells, located at the surface ofthe cellular aggregates, were small, rounded, highly cytoplasmic,and rich in rough endoplasmic reticulum. Further within theaggregates there occurred some cells showing abnormal or incompletecytokinesis and having irregularly thickened walls. Locatedcentrally in the aggregates were cells showing massive accumulationsof electron-dense material and with cell walls showing bandsof thickening alternating with thinner wall regions traversedby plasmodesmata. The latter cells are interpreted as cellsshowing intense polyphenol metabolism and imperfect xylogenicdifferentiation.     

Internal Root Anatomy of Maize Seedlings (Zea mays L.) as Influenced by Temperature and Genotype

The objectives of this investigation were to determine: (a)the general effect of temperature on internal root anatomy;(b) whether genotypic differences in such root traits exist;and (c) the association between internal root traits and shootgrowth, lateral root branching and cold tolerance of maize (Zeamays L.). Seedlings of 20 central European hybrids were grownunder high or low temperature (25/22·5 °C or 15/12·5°C day/night temperatures) until the third leaf was fullyexpanded. Light microscopy of cross sections revealed a largerdiameter of primary roots at low temperature which was due toa larger stele diameter and a thickening of the cortex. Concurrently,an increase in total cross sectional area of metaxylem elementswas obtained. It is assumed that the modification of the internalroot structure by temperature has an effect on both axial andradial water flow capacity. For all anatomical traits studied,variability between genotypes was apparent under both growingconditions. Furthermore, different genotypic responses to temperaturewere observed. However, basic differences between cold-tolerantand cold-sensitive genotypes did not exist. While at high temperatureroot traits and shoot growth were significantly and positivelycorrelated, at low temperature the correlation coefficient wasinsignificant. Consequently, it was not possible to characterizethe performance of the shoot at low temperature based on anatomicaltraits of the root. Moderate, positive correlation coefficientswere obtained between internal root traits and lateral rootbranching. The potential use of root anatomical traits as indirectselection criteria is discussed. Chilling tolerance, genotypes, root anatomy, Zea mays L