Non-lignified helical cell wall thickenings in root cortical cells of Aspleniaceae (Polypodiales): histology and taxonomical significance

Background and Aims

Extraxylary helical cell wall thickenings in vascular plants are not well documented, except for those in orchid velamen tissues which have been studied extensively. Reports on their occurrence in ferns exist, but detailed information is missing. The aim of this study is to focus on the broad patterns of structure and composition and to study the taxonomic occurrence of helical cell wall thickenings in the fern family Aspleniaceae.

Methods

Structural and compositional aspects of roots have been examined by means of light, electron, epifluorescence and laser scanning confocal microscopy. To assess the taxonomical distribution of helical cell wall thickenings a molecular phylogenetic analysis based on rbcL sequences of 64 taxa was performed.

Key Results

The helical cell wall thickenings of all examined species showed considerable uniformity of design. The pattern consists of helical, regularly bifurcating and anastomosing strands. Compositionally, the cell wall thickenings were found to be rich in homogalacturonan, cellulose, mannan and xyloglucan. Thioacidolysis confirmed our negative phloroglucinol staining tests, demonstrating the absence of lignins in the root cortex. All taxa with helical cell wall thickenings formed a monophyletic group supported by a 100 % bootstrap value and composed of mainly epiphytic species.

Conclusions

This is the first report of non-lignified pectin-rich secondary cell walls in ferns. Based on our molecular analysis, we reject the hypothesis of parallel evolution of helical cell wall thickenings in Aspleniaceae. Helical cell wall thickenings can mechanically stabilize the cortex tissue, allowing maximal uptake of water and nutrients during rainfall events. In addition, it can also act as a boundary layer increasing the diffusive pathway towards the atmosphere, preventing desiccation of the stele of epiphytic growing species.     

Vestured pits and vestured vessel member walls in some Indian dicotyledonous woods

NAIR, M. N. B. AND MOHAN RAM, H. Y., 1989. Vestured pits and vestured vessel member walls in some Indian dicotyledonous woods. The woods of 144 taxa belonging to 38 families of angiosperms were examined for vestured pits and vestured vessel member walls using scanning electron microscopy. Vestured pits are present in 66 taxa (belonging to ten families) and vestured vessel member walls occur in only six taxa (belonging to three families). In Ehretiaceae and Euphorbiaceae vestures are present only in certain vessel members. In Wrighlia tinctoria , perforation plates containing vestures have been observed in addition to the presence of vestured pits. A classification of vestured pits based on their morphology and distribution is proposed by us. In all the types of vestured pits, vestures are present on the margin of the outer pit aperture or on the pit chamber wall. Occasionally, they are present in the pit canal, on the margin and in the vicinity of the inner pit aperture and rarely over the inner walls of the vessel members. The functions of vestured pits are not clear, although several suggestions are made. Whether or not these structures affect wood processing is not presently understood. It appears that vestured pits and vestured vessel member walls have diagnostic rather than phylogenetic value.     

Regulation of cell division in pea root tips after wounding: A possible role for ethylene

Summary Calcofluor White ST is a fluorescent brightener that has previously been shown to alter cellulose ribbon assembly in the bacteriumAcetobacter xylinum. In this report, we demonstrate that Calcofluor also disrupts cell wall assembly in the eukaryotic algaOocystis apiculata. When observed with polarization microscopy, walls altered by Calcofluor show reduced birefringence relative to controls. Electron microscopy has shown that these altered walls contain regions which consist primarily of amorphous material and which generally lack organized microfibrils. We propose that wall alteration occurs because Calcofluor binds with the glucan chains polymerized by the cellulose synthesizing enzymes as they are produced. As a consequence, the glucan chains are prevented from co-crystallizing to form microfibrils. Synthesis of normal walls resumes when Calcofluor is removed, which is consistent with our proposal that Calcofluor acts by direct physical interaction with newly synthesized wall components.Several types of fluorescent patterns at the cell wall/plasmalemma interface have also been observed following Calcofluor treatment. Fluorescent spots, striations; helical bands, and lens-shaped thickenings have been documented. Each of these patterns may be the result of the interaction of Calcofluor with cellulose at different spatial or temporal levels or from varying concentrations of the brightener itself. Helical bands and lens-shaped thickenings also have been examined with the electron microscope. Like other regions of wall alteration, they are found to contain primarily amorphous material. Finally, we note that cells with severely disrupted walls are unable to complete their normal life cycle.     

The anatomy of lotus fibers found in petioles of Nelumbo nucifera

Lotus fibers are the isolated helical secondary cell wall thickenings from tracheary elements of lotus (Nelumbo nucifera Gaertn) petioles. In this study the anatomical characteristics of lotus petioles and microstructures of tracheary elements were studied using light microscopy (LM) and scanning electron microscopy (SEM). The results show that vascular bundles of lotus petioles are scattered throughout ground tissue. Their tracheary elements are of various sizes and there are several patterns of secondary wall thickening present. However, only secondary thickening in a ribbon-like helical pattern can be drawn out from the petiole to form lotus fibers for subsequent utilization. Study of the microstructure of the tracheary elements reveals that there are two pit structures present in the end walls in addition to pits with intact pit membranes: those with porose or web-like remnants pit membrane and those that lack pit membranes. This is an indication of the transitional stage between tracheids and vessel elements. This study provides supportive evidence that lotus fibers are found in both helically thickened tracheids and helically thickened primitive vessels.     

Testing hypotheses that link wood anatomy to cavitation resistance and hydraulic conductivity in the genus Acer

? Vulnerability to cavitation and conductive efficiency depend on xylem anatomy. We tested a large range of structure-function hypotheses, some for the first time, within a single genus to minimize phylogenetic 'noise' and maximize detection of functionally relevant variation. ? This integrative study combined in-depth anatomical observations using light, scanning and transmission electron microscopy of seven Acer taxa, and compared these observations with empirical measures of xylem hydraulics. ? Our results reveal a 2 MPa range in species' mean cavitation pressure (MCP). MCP was strongly correlated with intervessel pit structure (membrane thickness and porosity, chamber depth), weakly correlated with pit number per vessel, and not related to pit area per vessel. At the tissue level, there was a strong correlation between MCP and mechanical strength parameters, and some of the first evidence is provided for the functional significance of vessel grouping and thickenings on inner vessel walls. In addition, a strong trade-off was observed between xylem-specific conductivity and MCP. Vessel length and intervessel wall characteristics were implicated in this safety-efficiency trade-off. ? Cavitation resistance and hydraulic conductivity in Acer appear to be controlled by a very complex interaction between tissue, vessel network and pit characteristics.     

Vestigial pit membrane remnants in perforation plates and helical thickenings in vessels of Ericaceae

Perforation plates from 15 species of 10 genera with scalarifom perforation plates, representing three subfamilies of woody Ericaceae (Rhododendroideae, Arbutoideae, Vaccinioideae) were studied with scanning electron microscopy (SEM). In most of them, pit membrane remnants were present, but these remnants were less extensive than in the ericalean families Clethraceae, Cyrillaceae, and Sarraceniaceae. Pit membrane remnants in perforations of vessels of Ericaceae are characteristically found at lateral ends of the perforations and in perforations (which may alternatively be called pits) transitional to lateral wall pitting. Pit membrane remnants were most extensive in Enkianthus. Phylogenetic and physiological factors for vestigial membrane presence in the perforations are discussed. Helical thickenings on vessel walls as seen with SEM are figured and described for Leucothoe and Pieris, and their significance is assessed.     

Can cell walls bending round xylem vessels control water flow?

Vascular bundles of petioles below wilted leaves of Nymphoides peltata (S.G. Gmel. O. Kuntze) were frozen intact and freeze-fractured for electron microscopy. Cell walls in them appeared drawn in against the helical thickenings of xylem vessels. By contrast, walls round vessels which had been frozen in vascular bundles below turgid leaves, and walls round vessels which had been fixed, embedded and sectioned, were straight or bulged outwards slightly. Walls bulged outwards slightly also from cut vessels filled with sucrose solution before freezing. Movement of vessel walls could produce the clicks audible when water cavitates in vessels, and might explain a variable resistance to the flow of water through plants.     

LIGHT AND ELECTRON MICROSCOPE STUDIES OF THE PRIMARY XYLEM OF RICINUS COMMUNIS

Scott , Flora Murray , Virginia Sjaholm, and Edwin Bowler (U. California, Los Angeles.) Light and electron microscope studies of the primary xylem of Ricinus communis. Amer. Jour. Bot. 47(3) : 162-173. Illus. 1960.–The development of annular and spiral vessels in Ricinus communis has been examined under light and electron miscroscopes. Under the light microscope it is seen that spiral elements make up the bulk of the primary xylem. Pits and plasmodesmata are ubiquitous and are demonstrable in vertical and end walls. Plasmodesmata are evident in spiral thickenings. During tissue growth, intercellular spaces are formed between surrounding cells and developing vessels. These circum-vessel spaces are first lined with and later occluded by suberinlike substances. Traces of a material similar in microchemical reaction are laid down in the middle lamella. A suberin-like lining, termed in this paper the lipid lining, stainable with dimethylaminoazobenzene, occurs in mature living vessel elements. Innumerable minute fat-like droplets, refringent, and stainable with Sudan III, Sudan Black and also with osmic acid, occur in the outer cytoplasm and appear to be attached to the vessel lining by fine protoplasmic strands. They presumably are the source of the wall deposits. After the death of the protoplast, the vessel walls appear completely suberized. When contiguous cells are removed by treatment with I₂ki-H₂SO₄, their site and the site of the intercellular spaces remain marked by linear suberized ridges on the vessel wall. Annular and spiral thickenings arise as cellulose strands and begin to lignify only when the vessel reaches maximum diameter. In transverse section, the broken end of an extracted spiral thickening appears stratified. Under the electron microscope, pits and plasmodesmata are evident in procambial and in differentiating xylem elements in all walls. Annular and spiral thickening are distinguishable first as closely woven microfibrillar cellulose bands. As lignin is deposited in the microfibrillar mesh, the thickenings become dense to the electron beam. Irradiation with the full strength of the electron beam distinguishes between spiral thickenings in younger and older vessels. Older spirals remain apparently unchanged. Younger spirals instantly swell, volatilize in part, and assume a moniliform outline. The bead-like swellings consist of a matrix partly transparent to the electron beam and an internal framework of a material comparatively dense to the electron beam. Similar intense irradiation differentiates between younger and older vessel linings. Older linings appear unchanged, while the younger react violently, volatilize in part and stabilize as an irregular coagulum set in a basic mesh. The volatilized substances appear as granules on lining surface or on substrate. The changing microfibrillar pattern of the cell wall is observed from the procambial stage to the final deposition of the lipid vessel lining.     

SEM studies on vessels in ferns. 2. Pteridium

Xylem from roots and rhizomes of two infraspecific taxa of Pteridium aquilinum was studied by means of scanning electron microscopy (SEM). All tracheary elements proved to be vessels. End wall perforation plates were all scalariform, lacked pit membrane remnants in at least the central part of the perforation plate, and varied with respect to width of bars, from wide to tenuous, and with respect to presence of pit membrane remnants. In addition, porose pit membranes on walls that are likely all lateral vessel-to-vessel walls must be considered to be perforations also, although different from those on end walls. Lateral wall perforation plates, hypothesized by one worker on the basis of tylosis presence but denied by another on the basis of light microscopy, were confirmed by demonstration of pores with SEM. In addition, lateral walls of Pteridium vessels bear some grooves interconnecting pit apertures; this feature is newly figured by SEM for ferns. Lateral wall pitting that is not porose may either have striate thickenings of the primary wall or be smooth. Vessel presence and degree of specialization in Pteridium vessels may bear a relationship to the wide ecological tolerances of the genus.     

Wood anatomy of Elaeagnaceae, with comments on vestured pits, helical thickenings, and systematic relationships

The secondary xylem of Elaeagnus, Hippophae, and Shepherdia is described and illustrated in detail. Shrubs and small trees of Elaeagnaceae have ring-porous or semi-ring-porous wood with simple perforation plates, vascular tracheids, fiber-tracheids, diffuse or rarely paratracheal axial parenchyma, and uni- or biseriate rays in Hippophae and Shepherdia, but wider rays in Elaeagnus. Walls of vessel elements, especially narrow ones, tracheids, or fiber-tracheids sometimes show helical thickenings; in a few instances these intergrade with small bud-like protrusions associated with pits. Scanning electron microscopy illustrates that small to vestigial vestures are present in all species studied, although nonvestured pits are also common. The analogous nature of vestures and helical thickenings is considered. Comparative wood anatomy suggests a rather isolated position of the family Elaeagnaceae; affinities with Rhamnaceae, Proteaceae, and Thymelaeaceae are discussed.     

Wound-induced and naturally occurring regenerative differentiation of xylem in Zea mays L.

The regenerative differentiation of xylem, both around a wound in the stem and at the root junction was studied in seedlings of maize. The regeneration of vessels around a wound was very small, being limited to the very young internodes and sharply declining basipetally. There were more regenerative vessel elements and they differentiated faster above the wound than below it. The regenerative vessel elements around the wound were characterized by helical or annular pattern of secondary wall thickenings. Wounding also resulted in the development of additional vascular anastomoses in the leaf immediately above the wound, and in differentiation of discontinuous vessels in adjacent bundles. Regenerative vessel elements were very common where the adventitious roots connected with the stem internodes, and exhibited pitted or reticulated secondary wall thickenings.     

Vessel origins in Nymphaeaceae: Euryale and Victoria

Metaxylem tracheary elements of roots have differentiation between end walls and lateral walls in both Euryale and Victoria End walls have narrower, more closely spaced bars and scalariform plates. primary walls of end walls (and, to a lesser extent, lateral walls) have striations that are thickened primary wall portions orientated in an axial direction. These striations are less common in Victoria than in Euryale. Although secondary wall strands between perforations occur in some dicotyledons, the report of primary wall striations is new; these can be seen with scanning electron microscopy (SEM) but not with light microscopy. Perforations occur irregularly and sometimes sparsely on end walls of tracheary elements of Victoria , but perforations were not observed in Euryale. Thus, Euryale satisfies one criterion for the presence of vessel (end wall different from lateral wall), whereas Barclaya satisfies another (perforations in end walls) and Victoria satisfies both. Vessel origins in Nymphaeaceae are important in illustrating that there may be multiple vessel origins in dicotyledons.     

Ultrastructure of stomatal development in Arabidopsis (Brassicaceae) leaves

Stomatal development was studied in wild-type Arabidopsis leaves using light and electron microscopy. Development involves three successive types of stomatal precursor cells: meristemoid mother cells, meristemoids, and guard mother cells (GMCs). The first two types divide asymmetrically, whereas GMCs divide symmetrically. Analysis of cell wall patterns indicates that meristemoids can divide asymmetrically a variable number of times. Before meristemoid division, the nucleus and a preprophase band of microtubules become located on one side of the cell, and the vacuole on the other. Meristemoids are often triangular in shape and have evenly thickened walls. GMCs can be detected by their roughly oval shape, increased starch accumulation, and wall thickenings on opposite ends of the cells. Because these features are also found in developing stomata, stomatal differentiation begins in GMCs. The wall thickenings mark the division site in the GMC since they overlie a preprophase band of microtubules and occur where the cell plate fuses with the parent cell wall. Stomatal differentiation in Arabidopsis resembles that of other genera with kidney-shaped guard cells. This identification of stages in stomatal development in wild-type Arabidopsis provides a foundation for the analysis of relevant genes and of mutants defective in stomatal patterning, cell specification, and differentiation.     

RETICULATE THICKENINGS IN SOME SPECIES OF JUGLANS

Reticulate thickenings in Juglans nigra, J. major, J. microcarpa, and J. californica (including J. hindsii) appear, under the transmitted light microscope, to be a cluster of very large or “gash-like” vessel-ray pits. The use of scanning electron microscopy shows that these so-called pits are thickenings on the inside wall of the vessel.     

Transfer cells and flange cells in sinkers of the mistletoePhoradendron macrophyllum (Viscaceae), and their novel combination

Summary The unusual thick-walled cells in contact with host and parasite vessels, first noted by Calvin 1967 in sinkers (structures composed of tracheary elements and parenchyma that originate from parasite bark strands that grow centripetally to the host vascular cambium and become embedded by successive development of xylem) of the mistletoePhoradendron macrophyllum (Englem.) Cockerell, have been investigated by modern methods of microscopy. The wall is thickest in cells abutting large-diameter host vessels, less so against smaller host vessels and those abutting sinker vessels. Transmission electron microscopy reveals the wall to be complex, consisting of a basement primary wall, upon which two developments of secondary-wall material occur. These are represented by lignified thickenings, in the form of flanges, and a labyrinth of wall ingrowths characteristic of a transfer cell. The wall ingrowths occur mostly in the primary-wall regions between the flanges, but when in contact with a large host vessel the ingrowths also differentiate on top of the flanges. Cells with such a transfer cell labyrinth have not been previously reported in the endophytic system of a mistletoe. The cells are confined to the xylary portion of the primary haustorium and sinkers. InP. macrophyllum, however, the cells differ from ordinary transfer cells in that they have differentiated as part of a flange parenchyma cell. This arrangement represents a novel anatomical situation. The name flange-walled transfer cell is used for these cells. The xylem of primary haustorium and sinkers also contain numerous ordinary flange cells. In both flange-walled transfer cells and ordinary flange cells the flanges are lignified and form a reticulate pattern of thickenings, separated by rounded areas of primary pit fields. The extent of development of the flange wall can vary in different parts of a sinker. At the host interface, the existence of a flange-walled transfer cell in direct contact with a vessel reflects a site associated with high loading into the parasite. Similarly, a labyrinth against a sinker vessel indicates a site of unloading from surrounding sinker tissue into the vessel for subsequent longdistance transport within the parasite.Dedicated to the memory of Dr. Katherine Esau (1898–1997)     

Water transport in xylem conduits with ring thickenings

Helical or annular wall thickenings are not only present in protoxylem, but may also he a feature of the tracheids or vessel elements of secondary wood. The frequency of their occurrence tends to be a function of climatic factors and conduit diameter. In order to obtain a functional explanation for these structures, the hydrodynamic behaviour of xylem conduits with various patterns of annular wall thickenings was investigated numerically using a commercial CFD (Computational Fluid Dynamics) package. The fluid flow phenomena are presented in detail. The calculations show that the developing pressure gradient of the structures with corrugated walls is in each case lower than that of a smooth pipe with a diameter corresponding to the distance between two opposite thickenings. Furthermore, complex flow patterns with circulation zones between the thickenings develop which are dependent on the geometry of the wall. It may be hypothesized that these circulation zones influence the lateral water flow. The results are discussed with regard to the relationships between the water conduction function of the xylem and ecological factors.     

The tracheid-vessel element transition in angiosperms involves multiple independent features: cladistic consequences

Current definitions of tracheids and vessel elements are overly simple. These definitions are based on light microscope studies and have not incorporated information gained with scanning electron microscopy (SEM) or transmission electron microscopy (TEM). Current definitions are based primarily on angiosperms, especially eudicots, and were devised before many basal angiosperms were carefully studied. When all sources of information are taken into account, one can recognize changes in six characters in the evolution of tracheids into vessel elements in angiosperms (or vice versa) as well as in other groups of vascular plants. There is an appreciable number of taxa in which all criteria for vessel origin are not met, and thus incipient vessels are present. At the very least, vessel presence or absence should not be treated as a single binary character state change in construction of cladistic matrices. Increase in conductive area of an end wall by means of lysis of progressively greater areas of pit membrane and increase in pit area on the end wall (as compared to pit area on equivalent portions of lateral walls) are considered the most important usable criteria for recognizing intermediacy between tracheids and vessel elements. Primitive character states in vessel elements are briefly discussed to differentiate them from changes in character states that can be regarded as intermediate between tracheids and vessel elements.     

The role of microtubules in vessel member differentiation inColeus

Summary The role of microtubules in tracheary element formation in cultured stem segments ofColeus has been investigated through the use of the antimicrotubule drug, colchicine. Colchicine treatment of the cultured stem segments produced a dual effect on xylem differentiation. If applied at the time of stem segment isolation or shortly thereafter, wound vessel member formation is almost completely blocked. However, if colchicine is applied after the third day of culture, it does not inhibit differentiation, but instead large numbers of xylem elements are formed which have highly deformed secondary walls. Both effects are related to colchicine's specific affinity for microtubules. In the first case it is shown that colchicine blocks mitosis, presumably by destroying the spindle apparatus, and thus inhibits divisions which are prerequisite for the initiation of xylem differentiation. While, if colchicine is applied after the necessary preparative divisions have taken place, it destroys specifically the cortical microtubules associated with the developing bands of secondary wall, thus causing aberrant wall deposition.Light and electron microscopic analysis of drug-treated cells reveals that the secondary wall becomes smeared over the surface of the primary wall and does not retain the discrete banded pattern characteristic of secondary thickenings in untreated cells. Examination of colchicine-treated secondary walls in KMnO₄ fixed material shows that in the absence of microtubules the cellulose microfibrils lose their normal parallel orientation and are deposited in swirls and curved configurations, and often lie at sharp angles to the axis of the secondary wall band. Microtubules, thus, appear to play a major role in defining the pattern of secondary wall deposition and in directing the orientation of the cellulose microfibrils of the wall. Factors in addition to microtubules also act in controlling the secondary wall pattern, since we observe that even in the absence of microtubules secondary thickenings of two adjacent xylem elements are deposited directly opposite one another across the common primary wall.     

10 种茶藨子属植物导管分子形态特征及其生态适应性比较研究简

利用光学显微镜和扫描电子显微电镜,以10种茶藨子属植物为研究材料,观察了其导管分子的形态结构。结果显示：（1）所研究的茶藨子属植物导管分子穿孔板为梯状穿孔板,且穿孔板上横隔分布的数量不同,端壁倾斜角度种间变化不大;（2）导管分子纹孔式样为互列式或兼有互列式和对列式,纹孔形状种间存在差异;（3）有的种类导管分子内壁有螺纹加厚或网状凸起。研究表明,不同生境下的茶藨子属植物导管形态与其生态适应性之间有较强的相关性,表现为湿生环境的物种导管较短,直径较宽;旱生环境的物种导管较长,直径较小;中生环境的居中。本文分析了茶藨子属植物导管分子形态特征的生态适应性。