The effects of lead,nickel, and strontium nitrates on cell division and elongation in maize roots

The effects of Pb, Sr, and Ni nitrates on the root growth, its cell division and elongation were studied. Two-day-old maize seedlings were incubated on the 35 μM Ni(NO₃)₂, 10 μM Pb(NO₃)₂, or 3 mM Sr(NO₃)₂ in the presence or absence of 3 mM Ca(NO₃)₂. Metal toxicity was evaluated after the inhibition of root growth for the first and second days of incubation in comparison with the roots kept on water or Ca(NO₃)₂ solution. The contents of metals were determined in the apical (the first centimeter from the tip) and basal (the third centimeter from the kernel) root parts by voltamperometry and atomic-absorption spectrophotometry. We measured the length of the meristem, the length of the fully elongated cells, counted the mitotic index (MI) in the meristem and the number of meristematic cells in the cortex row; we also calculated duration the cell cycle. In the absence of Ca(NO₃)₂, the metal content in the apical root region was higher than in basal one. In the presence of Ca(NO₃)₂, we observed reverse ratio most pronounced in the case of Pb and Sr. All metals tested markedly reduced MI in the cortex, which was determined by the increase in the cell cycle duration and accompanied by the meristem shortening. These metals affected differently cell division and elongation: Ni inhibited mainly cell division and to a lesser degree their elongation, whereas Sr and Pb affected both cell division and elongation; only Sr treatment resulted in the increased length of the fully elongated cells. In the presence of Ca, all studied growth indices changed less than in the absence of Ca, which was manifested in the less severe suppression of the root growth and was in agreement with the lower accumulation of the metals in the root tips. Possible causes for the heavy metal action on growth are discussed in connection with the specificity of their transport and accumulation.     

Enhancement of nickel and lead accumulation and their toxic growth-inhibitory effects on amaranth seedlings in the presence of calcium

The effects of Ni(NO₃)₂ and Pb(NO₃)₂ on Amaranthus sp. L. root growth and the effect of calcium on heavy metal (HM) accumulation in the growing root zone and root growth inhibition were studied. The seeds were germinated in the Pb(NO₃)₂ solutions at concentrations of 50, 100, 200, 500 and 700 μM or Ni(NO₃)₂ solutions at concentrations of 10, 50, 70, 100, and 500 μM in the presence of 100 μM Ca(NO₃)₂ or without it. HM toxicity was assesses in 7 days after seed sowing by the root length. Distribution of HM over the tissues of the growing root part was examined histochemically. Ni was more toxic to root growth than Pb. In the presence of Ca, Ni and Pb accumulation in the amaranth root growing part increased markedly, and this enhanced their growth-inhibitory of action. A comparison of results obtained in this work and available from the literature permitted a conclusion that the routes of HM penetration into the root differ in different plant species, and this determines ambiguity of protective Ca action.     

Stem cells in the root and the problem of stem cells in plants

Plant cells are capable of reversible transition from the proliferating to the stem state. This transition is determined by a system of cell-cell interactions and interelationships between plant parts. Stem cells defined as the cells preserving the capacity to divisions and differentiation for a long time arise repeatedly during development of the root and shoot primordial, rather than are clones of a population of stem cells laid down at a certain stage of embryogenesis. The quiescent center cells, rather than the surrounding actively dividing cells, best correspond to the characteristics of stem cells according to Loeffler and Potten. The factors that determine the quiescent center formation and maintenance in the root have been analyzed. The available data suggest that among these factors, indoleacetic acid transport and cap influence are of paramount significance. The cap formation precedes the quiescent center formation both during the root development and in the course of meristem regeneration after the root decapitation. The capacity of stem cell formation by the meristem suggests that not only meristem arises from the stem cells, but also that stem cells are formed from actively dividing cells. Repeated formation of stem cells allows long-term preservation of the capacity of plants for open morphogenesis and vegetative propagation.     

Three maize root-specific genes are not correctly expressed in regenerated caps in the absence of the quiescent center

The quiescent center is viewed as an architectural template in the root apical meristem of all angiosperm and gymnosperm root tips. In roots of Arabidopsis thaliana (L.) Heynh., the quiescent center inhibits differentiation of contacting initial cells and maintains the surrounding initial cells as stem cells. Here, the role of the quiescent center in the development of the maize (Zea mays L.) root cap has been further explored. Three maize root-specific genes were identified. Two of these were exclusively expressed in the root cap and one of them encoded a GDP-mannose-4,6-dehydratase. Most likely these two genes are structural, tissue-specific markers of the cap. The third gene, a putative glycine-rich cell wall protein, was expressed in the cap and in the root epidermis and, conceivably is a positional marker of the cap. Microsurgical and molecular data indicate that the quiescent center and cap initials may regulate the positional and structural expression of these genes in the cap and thereby control root cap development. Received: 22 September 1999 / Accepted: 9 November 1999     

Effect of nickel at high concentration on proliferation of quiescent center cells and initiation of lateral root primordia in wheat seedlings

DNA synthesis and cell divisions in the quiescent center as well as initiation of lateral root primordia were investigated in the course of incubation of the roots of 3-day-old wheat (Triticum aestivum L.) seedlings on the medium with 0.1 mM NiSO₄ for 72 h. It was found that the earliest effect of nickel on proliferation of the quiescent center cells was associated with an increase in the mitotic index 6 h after the beginning of its action. This effect was assumed to depend on an increase in mitosis time. Twelve hours after the beginning of the effect of nickel, mitotic index became somewhat lower, and in 18 h it sharply decreased. Some dividing cells were observed among the initial cells of certain tissues and near the quiescent center even in 72 h. The portion of DNA synthesizing cell sharply decreased in 12 h, and in 48 h such cells were lacking. The main mechanism governing the termination of cell proliferation in the quiescent center as well as in the meristem and calyptrogen of the cap is the inhibition of cell transition to DNA synthesis. The cells that had time to start DNA synthesis or already finished it and were in other phases of the cycle continued a slow progression through the cycle and completed it. Sister cells, produced as a result of divisions, left the mitotic cycle in the phase G₁ and transited to dormancy. Nickel did not inhibit initiation and development of lateral root primordia. Resumption of DNA synthesis and cell divisions occurred not only in the pericycle and endodermis participating in the initiation of lateral root primordia but also in the cortex cells in the vicinity of developing primordia. In 18 h after the beginning of the experiment when the rate of the root growth considerably decreased, the region, where primordia were initiated, was located closer to the root tip. Subsequently, when elongation of the cells was inhibited, this region moved closer to the tip until structural disturbances occurred in the nuclei of the endodermal cells located near the root tip and elongated under the effect of nickel. The results concerning the effect of nickel and other heavy metals on root cell proliferation obtained by other researchers and the role of pericycle organization in the translocation and accumulation of nickel in the tissues are discussed.     

Nickel Toxicity and Distribution in Maize Roots

A new histochemical method for Ni determination has been developed and employed to study the pattern of Ni distribution in plant tissues. Two-day-old seedlings of maize (Zea mays L.) were transferred onto 15, 20, 25, and 35 M Ni(NO₃)₂ solutions in the presence of 3 mM Ca(NO₃)₂, and Ni localization in shoot and root tissues was investigated at days 2 and 7 of the incubation. Following two days of incubation, Ni was found in all root tissues, and its content increased with the period of exposure and from the tip to the root base. Independent of root region and tissue, Ni content in the protoplasts exceeded that in the cell walls. Ni penetrated the endodermal barrier and accumulated in the endodermis and pericycle to the highest concentration. Ni accumulation in the pericycle restricted root branching. Ni did not affect the final cell length, and the inhibition of root growth resulted from suppressed cell division. In the shoots, Ni content was below the level discerned by the dimethylglyoximine method; we therefore conclude that maize belongs to excluder plants, with their root systems functioning as a barrier limiting heavy metal intake by aboveground organs. The pattern of Ni transport differs from that of Cd and Pb; this difference stands for specific toxic effects of Ni, including an arrest of root branching.     

Stem cells in the root and the problem of stem cells in plants

Plant cells are capable of reversible transition from the proliferating to the stem state. This transition is determined by a system of cell-cell interactions and interrelationships between plant parts. Stem cells defined as the cells preserving the capacity to divisions and differentiation for a long time arise repeatedly during development of the root and shoot primordial, rather than are clones of a population of stem cells laid down at a certain stage of embryogenesis. The quiescent center cells, rather than the surrounding actively dividing cells, best correspond to the characteristics of stem cells according to Loeffler and Potten. The factors that determine the quiescent center formation and maintenance in the root have been analyzed. The available data suggest that among these factors, indoleacetic acid transport and cap influence are of paramount significance. The cap formation precedes the quiescent center formation both during the root development and in the course of meristem regeneration after the root decapitation. The capacity of tem cell formation by the meristem suggests that not only meristem arises from the stem cells, but also that stem cells are formed from actively dividing cells. Repeated formation of stem cells allows long-term preservation of the capacity of plants for open morphogenesis and vegetative propagation.     

The Difference Between Open and Closed Meristems

An open and a closed root meristem have been compared by investigatingthe cell kinetics of small regions of the apices of Helianthusand Zea. The cells of the stelar pole are quiescent in both and thereis no exchange of cells between stele and cortex or stele andcap. The immediately distal cells in the closed meristem (Zea)are also quiescent and the few divisions that do occur can betransverse or longitudinal. In the open meristem (Helianthus)these cells are not quiescent, but they go out of cycle transiently,prolonging the potential cell-doubling time. Their divisionsare transverse. It is a consequence of these differences thatclosed meristems form root caps discrete from the cortex whereasopen meristems force instability in the boundary between theperipheral part of the cap and the cortex. Another consequencein roots with open meristems is a succession of columella complexestransversely displaced from each other by the state of fluxin the meristem during the non-cycling phase of the proximaltier of cells, those immediately distal to the stelar pole. The results are discussed in relation to the ontogenetic onsetof quiescence and the evidence for switches between open andclosed operation of meristems. meristem, root apex, Helianthus annuus, Zea mays L.     

Effect of auxin on acropetal auxin transport in roots of corn

Acropetal [¹⁴C]indoleacetic acid (IAA) transport was investigated in roots of corn. At least 40 to 50% of this movement is dependent on activities in the root apex. Selective excision of various populations of cells comprising the root apex, e.g. the root cap, quiescent center, or proximal meristem show that the proximal meristem is the critical region in the apex with regard to influencing IAA movement. The quiescent center has no influence and the root cap has only a minor effect. Excision and replacement of the proximal meristem with an exogenous supply of 10⁻⁸ to 10⁻⁹ molar IAA prevents the reduction in acropetal IAA transport which would normally occur in the absence of this meristem. Substituting 10⁻⁹ molar IAA for the excised root cap brings about a significant increase in the amount of IAA moved acropetally, as compared to intact roots with the root cap still in place. From this and previous work, it is concluded that IAA synthesis occurring in the proximal meristem stimulates the movement of IAA from the basal to apical end of the root.     

The influence of lead on callose formation in roots ofLemna minor L

The treatment ofLemna minor L. plants with Pb(NO₃)₂ for 90 min, 8 and 24 h resulted in intensified deposition of (l,3)-P-glucan (callose) in plants roots. It was localized in the protodermis of the root tip, and in the center of the stele in the region at the proximal part of the root cap and slightly above     

In Vitro Transformation of Root meristem to Shoot and Plantlets in Vanilla planifolia

A study is reported of histogenesis and organogenesis duringthe processes leading up to plantlet formation in tip culturesof aerial roots of Vanilla planifolia. Young root tips excisedfrom aerial roots, less than 15 cm long, when cultured in liquidMS medium containing IAA and KN showed gravitropic responseuntil cap lysis began. With the collapse of the distal halfof the cap, the cells of the quiescent centre divided forminga hemispherical mass of cells. Further localized divisions onthe periphery of the hemisphere resulted in a number of meristemoidseach of which differentiated into a shoot meristem with leafprimordia. Procambium differentiated first beneath the apicalmeristem after two to three leaf primordia had formed and thenat the base of the leaves. After a few leaves have been formeda root meristem differentiated in close lateral proximity tothe basal end of the shoot procambium. Formation of a plateof vasculature at the nodal region of the first formed leaf,procambialization of the root and the bridging up of the shootand root vasculature with the nodal plate are described. Vanilla planifolia, root tip, in vitro, quiescent centre, meristemoid, plantlet     

STUDIES IN THE QUIESCENT ROOT MERISTEM OF THE BULB OF ALLIUM CEPA L.: FINE STRUCTURE AND DNA SYNTHESIS *

Study of the quiescent root meristem of Allium cepa L. bulbs has revealed that its histological organization does not differ significantly from the growing meristem, except for the fact that the cells are all arrested in interphase. Ultrastructure of the quiescent tissue is, however, different in the organization of the nucleolus and in the absence of prominent endoplasmic reticulum, microtubules and golgi complexes. A variety of lomasome-like structures, plasma membrane modifications and vacuoles have been recorded. Most of the cells except for the ones in the root cap and quiescent center are highly vacuolated; vacuolation is maximum in the cortical zone of the meristem. The pattern of ³H thymidine incorporation during early stages of sprouting indicates that asynchrony of the mitotic cycle, which is the characteristic of the growing meristem, is maintained during quiescence by the arrest of nuclei at different subphases of interphase.     

DURATION OF CELL CYCLES IN CULTURED ROOTS OF CONVOLVULUS

The durations of the cell cycle in physiologically different regions of the meristem of cultured roots of Convolvulus arvensis were determined by the metaphase-accumulation technique involving colchicine. The cell cycle in the root cap increases from 13 hr in the actively dividing initials of the first tier to 155 hr in the slowly dividing initials of tiers 2–4 to an indeterminate value for derivatives of the initials in the root cap columella. The cycle times for the cells of the central cylinder and cortex are 21 and 27 hr, respectively. The cells of the quiescent center have a cycle of an estimated 420 hr. The duration of the cell cycle in these different regions is discussed in relation to the increased duration of G₁ in slowly or non-dividing cells. The possible regulation of cell division by the synthesis of a cell-division factor in the quiescent center is also discussed.     

Magnetic field affects meristem activity and cell differentiation in Zea mays roots

Abstract

Exposure of Zea mays seedlings to a continuous electromagnetic field (EMF) for 30 h induced a 30% stimulation in the rate of root elongation compared with the controls. It also resulted in a significant increase of cell expansion, in both the acropetal (metaxylem cell lineage) and basipetal (root cap cells) direction. In addition, in EMF-exposed roots a precocious structural disorder was observed both in differentiating metaxylem cells and root cap cells. All these features may be consistent with an advanced differentiation of root cells that are programmed to die. EMF treatment also resulted in a significant reduction in the size of the quiescent centre in the root apical meristem. The extent to which these responses are causally linked is discussed.     

ROOT APEX STRUCTURE IN EPHEDRA MONOSPERMA AND EPHEDRA CHILENSIS (EPHEDRACEAE)

The root meristem of E. monosperma and E. chilensis possesses a central group of distinctive, large cells. These cells have large nuclei with scattered heterochromatin, proplastids with no starch, small vacuoles, mitochondria, few dictyosomes and endoplasmic reticulum cisternae, and lipid deposits. Over a 24 hr labelling period, the large cells fail to incorporate ³H-thymidine, whereas cells both distal and proximal to this region do. A quiescent center which includes these large cells is present therefore. Both species have an extensive root cap, the length being contributed by mitoses in many tiers of cells distal to the quiescent center. The root cap consists of a columella and peripheral regions. Distinctive amyloplasts, an increase in the number of endoplasmic reticulum cisternae and dictyosomes, large vacuoles, and lipid deposits are characteristic of differentiated columella cells. Peripheral cells elongate, lose most of their starch, and are eventually sloughed from the root.     

Influence of a weak DC electric field on root meristem architecture

BACKGROUND AND AIMS: Electric fields are an important environmental factor that can influence the development of plants organs. Such a field can either inhibit or stimulate root growth, and may also affect the direction of growth. Many developmental processes directly or indirectly depend upon the activity of the root apical meristem (RAM). The aim of this work was to examine the effects of a weak electric field on the organization of the RAM. METHODS: Roots of Zea mays seedlings, grown in liquid medium, were exposed to DC electric fields of different strengths from 0.5 to 1.5 V cm(-1), with a frequency of 50 Hz, for 3 h. The roots were sampled for anatomical observation immediately after the treatment, and after 24 and 48 h of further undisturbed growth. KEY RESULTS: DC fields of 1 and 1.5 V cm(-1) resulted in noticeable changes in the cellular pattern of the RAM. The electric field activated the quiescent centre (QC): the cells of the QC penetrated the root cap junction, disturbing the organization of the closed meristem and changing it temporarily into the open type. CONCLUSIONS: Even a weak electric field disturbs the pattern of cell divisions in plant root meristem. This in turn changes the global organization of the RAM. A field of slightly higher strength also damages root cap initials, terminating their division.     

Root apical organization inArabidopsis thaliana ecotype ‘WS’ and a comment on root cap structure

Arabidopsis thaliana roots have closed apical organization with three initial tiers. The dermatogen/calyptrogen tier consists of two parts-the central initials form the columella root cap, and the peripheral initial cells form the protoderm (epidermis) and the peripheral root cap. These peripheral initials divide in a sequence to form a root cap consisting of interconnected cones. the periblem initial tier forms the ground meristem (cortex). For the first week after germination the periblem consists of one layer of initial cells. The peripheral cells of the tier divide periclinally and then anticlinally (a T-division) to form the two-layered cortex (outer cortex and endodermis). After about one week, all the peripheral cells have divided periclinally forming two initials; the outermost produces the outer cortex while the inner initial produces the endodermis and middle cortex layer. The latter two cells arise via a periclinal division. During this time, other cells within the tier divide periclinally to form a two-layered tier. The plerome forms the cells of the procambium (vascular cylinder) by simple anticlinal divisions followed by longitudinal divisions to fill out the cell files of the vascular cylinder. A survey (27 dicot species in 17 families) of roots with closed apical organization revealed that there are three different types of root cap-concentric cylinders of cells (e.g.Linum), interconnecting cones (e.g.Arabidopsis) or overlapping arcs (e.g.Gossypium). H Lambers Section editor     

Evidence for the Active and Passive Chemotropisms in Roots

Changes in the elongation of root cells during the negative (away from the salt) or positive (towards the salt) chemotropic bending of roots induced by the unilateral application of agar blocks (1 mm³) with 10^–3and 10^–2M Cd(NO₃)₂solutions to the meristem zone of the root were studied. The root bending was not accompanied by differential changes in the number of cells that elongated during the 3-h period of chemical stimulation. The bending was only due to differential changes in the cell elongation rates. In most chemically stimulated roots, both concentrations of Cd(NO₃)₂solutions inhibited cell elongation at the stimulated and nonstimulated sides. Cell elongation was inhibited by 10^–2M Cd(NO₃)₂mainly on the stimulated side of the roots, hence, the roots bent towards the salt. On the contrary, 10^–3M Cd(NO₃)₂inhibited cell elongation mainly at the nonstimulated side of the roots. As a result, the roots bent away from the salt, i.e., in the direction opposite to that expected in the case of the direct inhibition of cell growth by Cd(NO₃)₂. It is concluded that the root chemotropisms induced by the above two Cd(NO₃)₂concentrations are, correspondingly, of a passive or active nature.     

Restoration of cell growth and proliferation in the wheat roots after their inhibition by nickel sulfate

The dynamics of cell growth and proliferation restoration in different tissues and quiescent center (QC) in the wheat (Triticum aestivum L.) seedling roots and also the differentiation of rhizodermal cells and lateral root initiation after 48-h treatment with 100 μM NiSO₄ were studied. Within 24 h after nickel removal from medium, root growth was resumed due to the increase in the rate of cell growth in the meristem and the region where cell elongation started in control roots. Stimulation of cell proliferation was restored in the main part of the meristem and later in the initial cells of the files and QC. Cell proliferation was not observed in the QC. The time of cell proliferation resumption in the roots and in tested tissues depended on the degree of their injury by nickel treatment. In most tested roots, DNA synthesis and cell division were restored in 32 h. In the cells leaving the meristem due to the resumption of their growth and proliferation, growth of root hairs started. In 48 h, the number of roots with perished cells in the rhizodermis in the meristem was sharply increased and the regeneration of the damaged region by the cells of outer cortex was observed. Only after the appearance of root hairs, the cells coming from the meristem started to elongate. In most roots, the formation of the new elongation zone occurred in 56 h. During its formation, the initiation of lateral root primordia was shifted in the basipetal direction. It was concluded that the cessation of cell growth and proliferation under the influence of high concentration of heavy metal (HM) ions is not lethal for the root. At the action of toxic HM concentrations, the plant strategy is the maintenance of meristematic cell capacity for cell growth and proliferation resumption. The cellular mechanism of this capacity maintenance is the transition of meristematic cells from G₁ phase to dormancy due to growth inhibition and the inhibition of the transition to DNA synthesis.     

The de novo origin of the quiescent center regenerating root apices of Zea mays

Summary In roots from which the root cap and quiescent center have been removed new apical tissues regenerated in line with the main axis of the root. Regeneration of these tissues occurred from the region of the proximal meristem, which extends for no more than 350 m from the cut surface. Accompanying the regeneration of new apical tissues is a change in the architecture of the root apex and initiation and enlargement of a new quiescent center. A possible role for the quiescent center in the establishment of pattern at the apex is considered. Regeneration of the original apex failed to occur in those roots from which the root cap, quiescent center and proximal meristem were excised.Abbreviation QC quiescent center