Development of cell wall modifications in the endodermis and exodermis of Allium cepa roots

The cytological events of wall modification in the endodermis and exodermis of Allium cepa L. roots were examined with fluorescence and transmission electron microscopy. In the endodermis, Casparian bands, suberin lamellae, and tertiary walls developed in succession. At the site of the future Casparian band, the plasma membrane was bound to the wall before deposition of detectable hydrophobic components in the radial wall. Suberin lamellae were deposited on the inner faces of the primary walls, first along the outer tangential walls and then the inner tangential walls. On both walls, segments of the lamellae were formed earlier in primary pit fields than at nonprimary pit field regions. Suberin lamellae then extended to the radial walls. When they reached the Casparian bands, the lamellae intruded between the bound plasma membranes and the walls, so that the cells' plasma membranes remained intact. In this way, suberin lamellae that were continuous around the cells were laid down. Later, tertiary walls were deposited internal to the suberin lamellae. None of the wall modifications interrupted the symplastic connections of the endodermis. During suberin lamella and tertiary wall formation, more dictyosomes and ER profiles appeared than during Casparian band development. In the exodermis, although Casparian bands were readily detected with fluorescence microscopy, they were rarely detected with electron microscopy. Suberin lamellae were formed in long cells severing their plasmodesmata. As in the endodermis, dictyosomes and ER were prominent during suberin lamella formation. Tertiary walls were not formed in the exodermis.Key words: Allium cepa, Casparian band, endodermis, exodermis, suberin lamella, ultrastructure.

Detecting exodermal Casparian bands in vivo and fluid-phase endocytosis in onion (Allium cepa L.) roots

Maturation of the exodermis involves development of a Casparian band, a structure that blocks the apoplastic movement of ions. The position at which this band is formed is not readily predictable, since it depends on species and growing conditions. Until now, Casparian band detection necessitated destructive methods which involved sectioning or clearing the roots. In the present study, a method for detecting exodermal Casparian bands in vivo is presented. Undisturbed onion roots were incubated for 2 h in 0.1% 8-hydroxy-1,3,6-pyrenetrisulphonate (PTS) and then thoroughly rinsed in running water. Under UV light, the tracer was evident in the cortex of the root regions with an immature exodermis but not in older regions of the root where the Casparian band had developed. PTS had entered the protoplasts of the cortical cells in the younger part of the root and had not been removed by rinsing. The first order kinetics of uptake, and insensitivity to external pH and probenecid indicated that PTS entered the cell by fluid-phase endocytosis. PTS-loaded vesicles that released their contents into the vacuole were seen using confocal laser scanning microscopy. When applied to undisturbed, whole root systems, PTS was not detected in aqueous extracts of the leaves. Thus, there is no major apoplastic bypass in healthy onion roots.Key words: exodermis, Casparian bands, apoplastic bypass, PTS, fluid-phase endocytosis.

Development of the endodermis and hypodermis of Typha glauca Godr. and Typha angustifolia L. roots

The development of the endodermis and hypodermis in adventitious roots of Typha angustifolia L. and Typha glauca Godr. was followed from the apical meristem to full maturity. The endodermis was typical, developing a thin Casparian band near the root tip, followed by suberin lamellae and asymmetric, secondary, lignified walls (C-type at maturity). Passage cells were present at an intermediate stage but eventually disappeared when all cells developed lamellae and secondary walls. The hypodermis was multiple (four to six layers at maturity) and began differentiating near the root tip. Here, the radial and transverse walls of the outermost layer did not dissolve in strong acid and the former were wavy in the longitudinal direction, both features characteristic of a Casparian band, but these walls were permeable to berberine. No other indication of a wall modification was seen for 3 weeks, at which time the root had become determinate and aerenchyma was beginning to form in the midcortex. Casparian bands, which were impermeable to berberine, matured in the hypodermis; thus, it proved to be an exodermis. Different forms of Casparian band were detected: one was typical and occupied the radial and transverse walls of the outermost layer, but others were novel and included tangential walls, often forming an H-shaped structure (as seen in cross section of the root). We propose calling the latter type an H-type Casparian band. It functioned as an apoplastic barrier to berberine applied either externally or internally by injection into the cortical aerenchyma. Following maturation of the Casparian band, the outer two layers of the exodermis soon produced suberin lamellae. These continued to be deposited in a centripetal pattern until they were found in all layers of the multiple exodermis. Development of the suberin wall modifications correlates with the development of the aerenchyma and may play a role in preventing gas exchange between the root and the rhizosphere. Later, all exodermal cells produced lignified, secondary walls. These were asymmetric in the outermost and innermost layers of the hypodermis (like the C-type endodermis); eventually, all layers had Casparian wall materials. Thus, the mature hypodermis consisted of two to six layers of exodermis, except at the tips of determinate roots where the exodermis was uniseriate with typical Casparian bands.Key words: cattail, endodermis, exodermis, hypodermis, roots, Typha.

Cortical ontogeny in roots of the aquatic plant, Hydrocharis morsus-ranae L.

Tissues in adventitious roots of Hydrocharis morsus-ranae L. developed from a four-tiered apical meristem. A set of periclinal divisions in the outermost layer of the ground meristem produced a hypodermis, which was normally uniformly biseriate. Aerenchyma formed from the adjacent inner layer of the cortex by a series of cell divisions and cell lyses; three- to five-celled, radial aerenchyma strands formed by periclinal divisions in radial cell files 0.3-5 mm behind the apex. Intervening cells underwent anticlinal and periclinal divisions followed by cell lyses within 1 mm of the apex to produce air spaces. Aerenchyma formation in this species is unusual and presents a unique system suitable for a study of developmentally programmed cell death in parenchyma cells. The endodermis formed a complete Casparian band about 10 mm behind the root apex and did not develop further; it had neither suberin lamellae nor secondary walls. The hypodermis was parenchymatous and was without Casparian bands, suberin lamellae, and secondary walls. Following acid digestion, the wavy walls of the endodermis and the walls of the epidermis remained.Key words: aerenchyma, cell death, endodermis, Hydrocharis, hypodermis, root development.

Suberin deposition and band plasmolysis in the corn (Zea mays L.) root exodermis

The exodermal Casparian band in corn (Zea mays L.) was first seen 10 mm distal to the kernel 4 days after planting. From its inception, the band usually occupied most of the radial wall (as seen in a cross section of the root). Subsequent maturation of the band around the root was asynchronous into the region of emerging lateral roots. Thus, a continuous apoplastic barrier would have been absent over much of the young root surface. Suberin lamellae development was also asynchronous, as these structures formed in those cells which had Casparian bands. Frequently, a lamella was initially deposited in patches, progressing centripetally until a continuous lipid layer was formed around the cell protoplast. Many instances of band plasmolysis (typical of the endodermis) were observed in the developing uniform exodermis. It could occur in cells with no detectable Casparian bands, suggesting that the tight connection between the plasmalemma and the wall that causes this phenomenon is not due to hydrophobic attractions. The results are consistent with the idea that there are strong attractions between proteins of the membrane and wall in the region of the Casparian band. The tight connection between the plasmalemma and the wall was broken during the later stages of suberin lamella development. Key words: Zea mays L., Poaceae, band plasmolysis, exodermis, Casparian band, suberin lamella.

Development of Casparian bands and suberin lamellae in the endodermis of onion roots

The endodermal Casparian band in onion (Allium cepa L.) roots is first evident 10 mm from the apex. It occupies a small region (20–30%) in the middle of the radial wall for a distance of 70 mm from the apex. However, with the development of suberin lamellae in most cells at greater distances from the root tip, the band expands symmetrically through the wall until, at 80 mm from the root tip, it fills the entire radial wall. Passage cells overlying some xylem poles have a delayed development of suberin lamellae, and in these cells the Casparian bands remain narrow. The sensitive stain, Fluorol yellow 088, detects precursors of the suberin lamellae in the outer tangential walls of the endodermal cells. The presence of complete lamellae is indicated by Sudan red 7B staining and resistance to acid digestion. Growth of the Casparian band during suberin lamella development may play a role in maintaining the apoplastic barrier in this layer by enlarging the contact area between the band and the lamellae. Key words: Casparian band, endodermis, suberin lamella, Allium cepa.

The apoplastic permeability of root apices

The apoplastic permeability of regions near the root tips of two nonexodermal species (Vicia faba L. and Pisum sativum L.) and three exodermal species (Zea mays L., Allium cepa L., and Helianthus annuus L.) was investigated with a berberine – thiocyanate tracer procedure. In regions of nonexodermal roots where the endodermis was mature, the walls of the epidermis and cortex were permeable; inward diffusion of the tracers was arrested at the endodermis. In regions of exodermal roots where the exodermis was mature, the apoplast was permeable only up to the anticlinal walls of the exodermis; in regions where the exodermis was immature but the endodermis was mature, the cortex was permeable up to the Casparian band of the endodermis. In root tips of both exodermal and nonexodermal species, berberine penetration into the meristem was greatly restricted. In some species there were additional areas adjacent to the meristem into which berberine movement was similarly restricted. It is concluded that even though root apices lack suberized structures, these zones are nevertheless partially apoplastically isolated from the external environment. Key words: apoplastic transport, berberine, fluorescent tracer, root tips.