The development of the endoder mis andPhi layer of apple roots

Summary Suberin lamellae and a tertiary cellulose wall in endodermal cells are deposited much closer to the tip of apple roots than of annual roots. Casparian strips and lignified thickenings differentiate in the anticlinal walls of all endodermal andphi layer cells respectively, 4–5 mm from the root tip. 16 mm from the root tip and only in the endodermis opposite the phloem poles, suberin lamellae are laid down on the inner surface of the cell walls, followed 35 mm from the root tip by an additional cellulosic layer. Coincidentally with this last development, the suberin and cellulose layers detach from the outer tangential walls and the cytoplasm fragments. 85 mm from the root tip the xylem pole endodermis (50% of the endodermis) develops similarly, but does not collapse. 100–150 mm from the root tip, the surface colour of the root changes from white to brown, a phellogen develops from the pericycle and sloughing of the cortex begins. A few secondary xylem elements are visible at this stage.Plasmodesmata traverse the suberin and cellulose layers of the endodermis, but their greater frequency in the outer tangential and radial walls of thephi layer when compared with the endodermis suggests that this layer may regulate the inflow of water and nutrients to the stele.     

Cluster root development in Grevillea robusta (Proteaceae). I. Xylem, pericycle, cortex, and epidermis development in a determinate root

The cluster roots of Grevillea robusta A. Cunn. ex R. Br. are composed of determinate rootlets that stop growing, but remain physiologically active for several months. Their apical organization, both before and after maturation, was studied by light and transmission electron microscopy. Each cell layer forms a dome, with an initial cell at its end. Xylem elements form a complicated triarch array at the base of the rootlet, passing along the rootlet as two files, and then joining at the tip to form a single file, surrounded by six pericycle cells. At the base of the rootlet, shorter xylem cells and thick-walled support cells are visible. A root cap, present in rootlets grown in vermiculite, was eventually displaced by root hair growth. Rootlets grown in Hoagland's solution lacked root caps and were significantly shorter than those grown in vermiculite. Cell fate was analysed in terms of cell position and is discussed in terms of pattern and development.     

Endodermal cell-cell contact is required for the spatial control of Casparian band development in Arabidopsis thaliana

Background and Aims

Apoplasmic barriers in plants fulfil important roles such as the control of apoplasmic movement of substances and the protection against invasion of pathogens. The aim of this study was to describe the development of apoplasmic barriers (Casparian bands and suberin lamellae) in endodermal cells of Arabidopsis thaliana primary root and during lateral root initiation.

Methods

Modifications of the endodermal cell walls in roots of wild-type Landsberg erecta (Ler) and mutants with defective endodermal development – scarecrow-3 (scr-3) and shortroot (shr) – of A. thaliana plants were characterized by light, fluorescent, confocal laser scanning, transmission and cryo-scanning electron microscopy.

Key Results

In wild-type plant roots Casparian bands initiate at approx. 1600 µm from the root cap junction and suberin lamellae first appear on the inner primary cell walls at approx. 7000–8000 µm from the root apex in the region of developing lateral root primordia. When a single cell replaces a pair of endodermal and cortical cells in the scr-3 mutant, Casparian band-like material is deposited ectopically at the junction between this ‘cortical’ cell and adjacent pericycle cells. Shr mutant roots with an undeveloped endodermis deposit Casparian band-like material in patches in the middle lamellae of cells of the vascular cylinder. Endodermal cells in the vicinity of developing lateral root primordia develop suberin lamellae earlier, and these are thicker, compared wih the neighbouring endodermal cells. Protruding primordia are protected by an endodermal pocket covered by suberin lamellae.

Conclusions

The data suggest that endodermal cell–cell contact is required for the spatial control of Casparian band development. Additionally, the endodermal cells form a collet (collar) of short cells covered by a thick suberin layer at the base of lateral root, which may serve as a barrier constituting a ‘safety zone’ protecting the vascular cylinder against uncontrolled movement of water, solutes or various pathogens.     

ADVENTITIOUS ROOT DEVELOPMENT FROM THE SEEDLING HYPOCOTYL OF LYCOPERSICON ESCULENTUM

Tomato seedlings five through ten days old were used for this investigation. Adventitious roots were initiated from the pericycle of the tomato hypocotyl. The position of adventitious root development was irregular in the rhizogenic hypocotyl; however, the cellular pattern of individual root development was very regular. Four layers of pericycle derivatives participated in root histogenesis and a bi- or triseriate endodermal cover was derived from the endodermis. Fluorescent microscopy showed that Casparian strips on the meristematic endodermal cell walls were not removed biochemically but were displaced around the root primordium by anticlinal divisions and cell enlargement. Casparian strips were not synthesized by endodermal cover cells. The emergent root had a typical three tiered or closed pattern of apical organization, and quiescent centers were present in all emergent roots longer than 0.5–0.6 cm.     

Ultrastructure and functioning of the transport system of the leguminous root nodule

Summary The structure of the vascular tissues of nitrogen-fixing nodules of 27 genera of legumes and some non-legumes has been investigated by light microscopy. Pisum and Trifolium nodules have been examined by electron microscopy.Attention is directed to the presence of a pericycle in the vascular bundles of the nodules. In 7 of the legumes the pericycle cells possess a wall labyrinth consisting of branched filiform protuberances. The ultrastructure of the pericycle cell cytoplasm is described: its most striking feature is its abundant rough endoplasmic reticulum. These cells surround the xylem and phloem of the bundles, and are in turn surrounded by a layer of endodermal cells with Casparian strips. The pericycle cells develop their wall labyrinth in the levels of the nodule at which the bacterial tissue becomes pigmented; in nodule senescence their cytoplasm is disrupted level with the breakdown of the bacterial tissue.A pathway for symplastic lateral transfer of assimilates exists, from the sieve elements through the pericycle, endodermis and cortex to the bacterial tissue. The apoplast within the endodermis consists largely of the pericycle wall labyrinth and the xylem. The ultrastructure of the Casparian strip resembles that of roots.Intact, detached nodules can be induced to bleed a fluid from their severed vascular tissue. This fluid is exceptionally rich in organic nitrogen, particularly amides, but does not appear to contain sugars. Comparison between its amino acid composition and that of other parts of the nodule suggests that an active uptake or secretion of nitrogenous compounds precedes export from the nodule. Special functions are suggested for the nodule endodermis and the pericycle cells in this export process.     

Comparative investigations on the differentiation of the endodermis and the development of the Hartig net in mycorrhizae of Picea abies and Larix decidua

Summary The differentiation of the endodermis of mycorrhizal roots of Picea abies and Larix decidua was investigated by means of light and transmission electron microscopy and with fluorescence techniques. The initiation and differentiation of the Hartig net were recorded. Differences between the two tree species were found, as were differences between the two tree species and angiosperms. The Casparian band developed immediately after the origin of endodermal cells from the meristem in mycorrhizae of both tree species. In L. decidua only the primary endodermis was present in most mycorrhizal laterals. The secondary structure of the endodermis was restricted to main roots and proximal parts of larch mycorrhizae. In P. abies mycorrhizae, however, the secondary stage of the endodermis developed soon after the primary endodermis and was characterized by regular alternation of short, active passage cells and elongated, rapidly degenerating cells, the inner surface of which was covered by a thick suberin layer. Hartig net development started in P. abies short roots only after the differentiation of endodermis into the secondary stage, whereas in L. decidua, the Hartig net was already initiated at the primary endodermal stage. Differences were specific for tree species.     

The Casparian Strip of Clivia miniata Reg. Roots: Isolation,Fine Structure and Chemical Nature*

The root endodermis of Clivia miniata Reg. was successfully isolated using the cell wall degrading enzymes cellulase and pectinase. The enzymes did not depolymerize those regions of the primary cell walls of anticlinal endodermal root cells where the Casparian strips were located. Since the endodermis of C. miniata roots remained in its primary developmental state over the whole root length, endodermal isolates essentially represented Casparian strips. Thus, sufficient amounts of isolated Casparian strips could be obtained to allow further detailed investigations of the isolates by microscopic, histochemical and analytical methods. Scanning electron microscopy revealed the reticular structure of the Casparian strips completely surrounding the central cylinder of the roots. Whereas in younger parts of the root only the anticlinal cell walls of the endodermis remained intact in the isolates, in older parts of the root the periclinal walls also restricted enzymatic degradation due to the deposition of lignin. Extracts of the isolates with organic solvents did not reveal any wax-like substances which might have been deposited within the cell wall forming a transport barrier, as is the case with cutin and suberin. However, several histochemical and analytical methods (elemental analysis and FTIR spectroscopy) showed that the chemical nature of the Casparian strips of C. miniata roots can definitely be a lignified cell wall. These findings are in complete agreement with studies carried out at the beginning of this century on the chemical nature of the Casparian strips of several other plant species. The implications of these results concerning apoplasmatic transport of solutes and water across Casparian strips are discussed.     

ORIGIN AND DEVELOPMENT OF LATERAL ROOTS IN TYPHA GLAUCA

Lateral roots of Typha glauca arose from the pericycle of the parent adventitious root. Periclinal divisions of the pericycle gave rise to two layers; the outermost initially produced the ground meristem and protoderm, and the innermost produced the procambium. The immature endodermis of the parent root contributed to the early stages of the root tip as an endodermal covering. Prior to emergence, the ground meristem/protoderm produced cells into the endodermal covering. After emergence, the endodermal covering was replaced by a calyptrogen, which was derived from the ground meristem/protoderm and which, in turn, formed the rootcap. A typical monocotyledonous three-tiered meristem was then produced. An outer ground meristem also arose before emergence to form a hypodermis in many lateral roots; in these, crystalliferous cell production began in midcortex cells before emergence, and a small aerenchyma developed in their cortices. The rootcap columella stored small amounts of starch shortly after emergence. Lateral roots of T. glauca were smaller than their parental adventitious roots; they normally had only two to six poles of xylem and phloem, and the cortex was less than six cells across. During 1–3-cm elongation, the lateral root apical meristem and mature regions narrowed, stored starch disappeared, fewer crystals formed, aerenchyma production ceased, and the roots stopped elongation.     

The Behaviour of Two Cell Populations in the Pericycle of Allium cepa, Pisum sativum, and Daucus carota During Early Lateral Root Development

The length of cells of the pericycle, endodermis and middlecortex not actively involved in lateral root primordia (LRP)development was measured in primary roots of Allium cepa, Pisumsativum and Daucus carota. The presence of two cell populationsin the pericycle was demonstrated in all three species. In Alliumcepa and Pisum sativum, pericyclic cells located opposite xylempoles were significantly shorter than cells lying opposite phloempoles. In both species, LRP originated opposite xylem poles.Our results, furthermore, strongly suggest that in regions ofthe root far from the apical meristem, numerous pericyclic cellsundergo transverse division both previous to and during LRPinitiation, decreasing in mean length throughout this period.In Daucus carota, LRP begin to form in pericyclic cells locatednext to the phloem poles, such cells were significantly shorterthan those opposite xylem poles, even in areas of the primaryroot located close to the root tip. Cells also appear to dividetransversely in regions far from the root tip in this species,leading to a conspicuous drop in the mean length of those cellslocated in portions of the pericycle destined to give rise toLRP. Two different cell populations can also be distinguishedin the endodermis of Allium cepa and Pisum sativum, althoughobservations were less conclusive in Daucus carota. In all threespecies, length of cortical cells was unaffected by their positionopposite xylem or phloem poles Allium cepa, carrot, cell division, cell length, Daucus carota, endodermis, lateral root development, onion, pea, pericycle, Pisum sativum     

Chemical composition and ultrastructure of broad bean (<Emphasis Type="Italic">Vicia faba</Emphasis> L.) nodule endodermis in comparison to the root endodermis

Ultrastructure and development of apoplastic barriers within indeterminate root nodules formed by Vicia faba L. were examined by light and electron microscopy. The nodule outer cortex is separated from the inner cortex by a heavily suberized nodule endodermis, which matures in submeristematic regions and possesses suberin lamellae. Unsuberized passage cells are present near vascular strands, which are surrounded by a vascular endodermis attached on the inner side of the nodule endodermal cell walls. The vascular endodermis appears immediately below the meristematic apex in developmental state I (Casparian bands), gradually develops suberin lamellae, and attains developmental state II at the base of the nodule. For chemical analysis apoplastic barrier tissues were dissected after enzymatic digestion of non-impregnated tissues. Root epidermal and endodermal cell walls as well as nodule outer cortex could be isolated as pure fractions; nodule endodermal cell walls could not be separated from vascular endodermal cell walls and enclosed xylem vessels. Gas chromatography-flame ionization detection and gas chromatography-mass spectrometry were applied for quantitative and qualitative analysis of suberin and lignin in isolated cell walls of these tissues. The suberin content of isolated endodermal cell walls of nodules was approximately twice that of the root endodermal cell walls. The suberin content of the nodule outer cortex and root epidermal cell walls was less than one-tenth of that of the nodule endodermal cell wall. Substantial amounts of lignin could only be found in the nodule endodermal cell wall fraction. Organic solvent extracts of the isolated tissues revealed long-chain aliphatic acids, steroids, and triterpenoid structures of the lupeol type. Surprisingly, extract from the outer cortex consisted of 89% triterpenoids whereas extracts from all other cell wall isolates contained not more than 16% total triterpenoids. The results of ultrastructural and chemical composition are in good correspondence and underline the important role of the examined tissues as apoplastic barriers.     

Effects of exposure to humid air on epidermal viability and suberin deposition in maize (Zea mays L.) roots

When the basal zones of 4-d-old hydroponically grown maize ( Zea mays L. cv. Seneca Horizon) roots were exposed to moist air for 2 d, the development of both endodermis and exodermis was affected. In the endodermis, Casparian bands enlarged and more cells developed suberin lamellae. The most striking effect was seen in the exodermis. In submerged controls, only 4% of the cells had Casparian bands, whereas in root regions exposed to air, 93% developed these structures. Similarly, in submerged roots 11% of the exodermal cells had either developing or mature suberin lamellae compared with 92% in the air-treated region. The majority of epidermal cells remained alive in the zone exposed to air. Some cell death had occurred earlier in the experiment when the seedlings were transferred from vermiculite to hydroponic culture. The precise stimulus(i) associated with the air treatment which led to accelerated development in both endodermis and exodermis is as yet unknown.     

Intercellular communication in the maize root endodermis

The endodermal cells of the maize roots are linked to each other via plasmodesmata mainly localized at the proximity of the Casparian band on the anticlinal walls. In this part of the walls was noticed the absence of the suberin lamella in the thickening endodermal cells.     

大豆初生根凯氏带对铜离子的通透性

采用在根内生成有色铜沉淀的方法研究大豆（Glycine max）初生根凯氏带对铜离子的通透性。用真空泵抽取浓度为200μmol·L^–1的CuSO4溶液进入根中,然后在重力作用下从根基部灌注400μmol·L^–1的K4[Fe（CN）6]溶液,两种物质在根内相遇即可产生棕色的Cu2[Fe（CN）6]沉淀,根据沉淀的位置来确定铜离子所经过的途径。结果表明：Cu^2＋可以穿过内皮层凯氏带,在木质部导管壁以及凯氏带至木质部之间的细胞壁处产生棕色沉淀,侧根发生的部位也产生了大量的沉淀;当抽取K4[Fe（CN）6]溶液后再灌注CuSO4溶液,发现Cu^2＋仍然可以穿过凯氏带,并在凯氏带外侧以及外皮层细胞的细胞壁处产生棕色沉淀。研究结果证明凯氏带并不是一个可以完全阻止离子进出的完美屏障。     

Root Endodermis and Exodermis: Structure, Function, and Responses to the Environment

Roots of virtually all vascular plants have an endodermis with a Casparian band, and the majority of angiosperm roots tested also have an exodermis with a Casparian band. Both the endodermis and exodermis may develop suberin lamellae and thick, tertiary walls. Each of these wall modifications has its own function(s). The endodermal Casparian band prevents the unimpeded movement of apoplastic substances into the stele and also prevents the backflow of ions that have moved into the stele symplastically and then were released into its apoplast. In roots with a mature exodermis, the barrier to apoplastic inflow of ions occurs near the root surface, but prevention of backflow of ions from the stele remains a function of the endodermis. The suberin lamellae protect against pathogen invasion and possibly root drying during times of stress. Tertiary walls of the endodermis and exodermis are believed to function in mechanical support of the root, but this idea remains to be tested. During stress, root growth rates decline, and the endodermis and exodermis develop closer to the root tip. In two cases, stress is known to induce the formation of an exodermis, and in several other cases to accelerate the development of both the exodermis and endodermis. The responses of the endodermis and exodermis to drought, exposure to moist air, flooding, salinity, ion deficiency, acidity, and mechanical impedance are discussed.     

Root Endodermis and Exodermis: Structure,Function, and Responses to the Environment

Roots of virtually all vascular plants have an endodermis with a Casparian band, and the majority of angiosperm roots tested also have an exodermis with a Casparian band. Both the endodermis and exodermis may develop suberin lamellae and thick, tertiary walls. Each of these wall modifications has its own function(s). The endodermal Casparian band prevents the unimpeded movement of apoplastic substances into the stele and also prevents the backflow of ions that have moved into the stele symplastically and then were released into its apoplast. In roots with a mature exodermis, the barrier to apoplastic inflow of ions occurs near the root surface, but prevention of backflow of ions from the stele remains a function of the endodermis. The suberin lamellae protect against pathogen invasion and possibly root drying during times of stress. Tertiary walls of the endodermis and exodermis are believed to function in mechanical support of the root, but this idea remains to be tested. During stress, root growth rates decline, and the endodermis and exodermis develop closer to the root tip. In two cases, stress is known to induce the formation of an exodermis, and in several other cases to accelerate the development of both the exodermis and endodermis. The responses of the endodermis and exodermis to drought, exposure to moist air, flooding, salinity, ion deficiency, acidity, and mechanical impedance are discussed.     

Relationships between structural development and the absorption of ions by the root system of Cucurbita pepo

Summary In both the seminal axis and lateral roots of Cucurbita pepo L. the formation of large central xylem elements and the commencement of secondary cambial activity occur 10–20 cm from the root tip. Concomitant with or slightly preceding these developments there are changes in the structure of the walls of endodermal cells where the lignified casparian band spreads along the radial wall and substances staining with Sudan IV are deposited in both radial and tangential walls. At distances more than 30 cm from the tip of primary roots the radius of the stele increases considerably causing splits in the cortex. The endodermis is stretched and the suberin becomes organized in a lamellar form.Against this background of anatomical change certain of the transport capabilities of the root are retained while others are lost. Using an apparatus for measuring the uptake of tracers by segments of intact roots it was found that neither the uptake nor translocation of potassium seem to be affected by the suberization of the endodermis or by secondary thickening, while the translocation of calcium is virtually eliminated when these processes begin. As the root ages its ability to absorb phosphate declines although the translocation of the phosphate absorbed is much less affected by structural development than that of calcium.The observed rates of potassium uptake by complete root systems could be predicted quite accurately from the average of segment uptake data suggesting that the method used gives reliable results.     

Chemical composition of Casparian strips isolated from Clivia miniata Reg. roots: evidence for lignin

Endodermal cell walls and xylem vessels were isolated enzymatically from Clivia miniata Reg. roots. Transmission-electron-microscopic investigation of cross-sections of intact C. miniata roots and scanning-electron-microscopic investigation of isolated endodermal cell walls indicated that the root endodermis of C. miniata is essentially in its primary state of development. Isolated Casparian strips and xylem vessels were subjected to two different degradation methods usually applied to prove the existence of lignin, namely, cupric oxide oxidation and thioacidolysis. The reaction products obtained were typical aromatic derivatives of the natural lignin precursors coniferyl and sinapyl alcohols, and, in traces, of p-coumaryl alcohol, indicating the occurrence of lignin in the polymers from both Casparian strips and xylem vessels. The qualitative chemical compositions of the polymers from the two sources were similar, whereas the quantitative compositions were different, indicating that the molecular structure of the lignin polymer in the Casparian strips was different from that in the xylem vessels. Thus, for the first time, direct chemical evidence has been obtained that Casparian strips of C. miniata roots contain lignin as a major cell wall polymer.The author is indebted to Prof. Dr. G. Krohne (Zentrale Abteilung für Elektronenmikroskopie, Universität Würzburg, Germany) and to Prof. Dr. R. Guggenheim (Labor für Rasterelektronenmikroskopie, Universität Basel, Schweiz) for offering the opportunity for transmission-electron-microscopic and low-temperature scanning-electron-microscopic investigations, respectively. Financial support by the Deutsche Forschungsgemeinschaft is gratefully acknowledged.     

Suberin lamella development in maize seedling roots grown in aerated and stagnant conditions

Hypoxia can stimulate the development of a suberized exodermis in aquatic plants; however, its influence on this aspect of terrestrial root development is sparsely documented. To determine the effects of hypoxia on maize (Zea mays cv. Seneca Horizon) roots, seedlings were grown in vermiculite (VERM), aerated hydroponics (AER), stagnant hydroponics with agar (STAG), or aerated hydroponics with agar (AERAG). The endo- and exodermis were examined for wall modifications. Lateral root emergence and aerenchyma formation were documented qualitatively. The endodermal Casparian band formation was unaffected by treatment. Endodermal and exodermal suberin lamella formation was earliest and most extensive in VERM. Suberization, especially in the exodermis of aerated treatments, was depressed in all hydroponic media. In comparison with AER, STAG exodermal lamellae were increased, but endodermal lamellae were decreased. Since the suberized exodermis forms a barrier to radial oxygen loss from roots to the medium, its stimulation in STAG roots (which also developed extensive aerenchyma) would help retain oxygen in the root. The reduction of endodermal lamellae should facilitate oxygen diffusion into the stele. Clearly, the response to environmental conditions is variable within individual cortical cell layers. Additionally, the observed patterns of lamellae, aerenchyma and lateral root development indicate a tight radial co-ordination of root development.