The Quiescent Centre and Rates of Mitosis in the Root Meristem of Allium sativum

The root apices of Allium sativum have been examined by continuous-and pulse-labelling with tritiated thymidine and by colchicinetreatment to measure the time parameters of the mitotic cyclein various parts of the meristem. There is a quiescent centre of 30–50 cells whose averagerate of mitosis is low because the G₁ period is extended toabout 140 h compared with about 4 h in the othe regions of themeristem. The stele just above the quiescent centre and at 200microns above it and the cap initials just below the quiescentcentre are very similar in their mitotic cycles, the total lengthsof which are about 30 h of which nearly half is taken up byDNA synthesis. Allium thus differs from Zea in having root capinitials whose mitotic cycle is not telescoped by the eliminationof the G₁ phase. These facts are discussed in relation to theradiosensitivity of the meristem.     

X-Irradiation of Root Meristems

When roots of Zea mays are given heavy doses of X-rays the averagerate of mitosis at first falls in the normally meristematiccells and increases in the quiescent centre. Proliferation inthe quiescent centre gives rise to a new meristem which continuesthe growth of the root and which acquires a quiescent centreof its own. Observations on the micronuclei produced after irradiationsuggest that the quiescent centre is less sensitive to X-raysthan the rest of the meristem. It is probably this lower sensitivitycombined with the initial difference in time at which the radiationdamage to cell viability occurs that gives the quiescent centreits role in the recovery of the meristem.     

Experimental control of the activity of the quiescent centre in excised root tips of Zea mays

Summary Quiescent centres have been demonstrated in cultured excised root tips of both Pisum sativum and Zea mays. Upon addition of sucrose to Zea roots which have been deprived of carbohydrate, the cells of the quiescent zone as well as those of the rest of the meristem undergo DNA synthesis. Following the onset of proliferative activity in the meristem, DNA synthesis in the quiescent-centre cells is again arrested. It is suggested that the dividing cells of the meristem are responsible for the maintenance of the quiescent centre. It has also been shown that DNA-synthesising cells do occur within the quiescent centre and that they appear to be localised in specific regions.     

The Response of the Primary Root Meristem of Zea mays L. to Various Periods of Cold

Primary roots of maize (Zea mays L.) grown in nutrient solutionat 5?C elongate at about 1% of the rate found at 20?C. The apicalmeristem becomes shorter and shows little proliferative activityat 5?C, but following transfer to 20?C mitoses increase in frequencyand the meristem regrows to its original length. Both the amountby which the meristem shortens and the time for its completeregrowth are related to the period spent at 5?C. The shorteningof the meristem suggests that at the lower temperature the balancewhich normally exists between cell production and differentiationis altered, the latter continuing at a relatively faster ratethan the former. A new, steady-state balance between the twoprocesses is re-established during the recovery period. Themeristem recovers as a result not only of its own mitotic activitybut also through stimulation of cell division in the quiescentcentre. The degree to which the quiescent centre is activated,as judged by its mitotic index and the number of nuclei labelledby feeding with tritiated thymidine, increases as the durationof the preceding cold treatment increases. The close relationshipbetween proliferative activity in the quiescent centre and theminimum length of the meristem following the cold treatmentsuggests that there is communication between these two zoneswhich co-ordinates their respective rates of cell productionand helps to maintain a normal meristem structure. The resultsemphasize the importance of the quiescent centre as a reservoirof cells that can re-establish a meristem rendered non-functionalthrough the impact of unfavourable environmental conditions. Key words: Meristem, quiescent centre, root, temperature, Zea mays     

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.     

Mitotic spindle and mitotic cell volumes in the root meristem of Zea mays

Summary Mitotic spindles in the root meristem of the Zea mays are smallest in the quiescent centre and increase in size the further they are from this region. the volume of mitotic cells follows a similar pattern. These findings are the result of differences in the metabolic activity of cells within the meristem. Observations also suggest that there may be fewer microtubules in the spindle of quiescent centre cells than in cells elsewhere, thus supporting the suggestion that this may be so made by Juniper and Barlow (1969).     

Nutrition and the quiescent centre of root meristems

Summary Root apices were fed with tritiated thymidine from the endosperm and from labelled root caps placed on decapped roots. Supply from both sources reaches the meristem, but is attenuated by the demand of the dividing cells. The thymidine reaches the far sides of the quiescent centre from both directions. Starvation is thus unlikely to be the reason for the existence of the quiescent centre although cessation of mitosis in the normally meristematic regions may stimulate the quiescent centre by releasing nutrients.     

Meristems under Continuous Irradiation

Root tips of Vicia faba and Zea mays have been subjected tocontinuous irradiation from radium or cobalt-60 for long periodsat various dose rates. The rates of mitosis and the damage tothe chromosomes (assessed as percentages of cells with micronulei)have been measured in four regions of the meristems. In Zearates of mitosis are reduced under chronic irradiation, exceptin the quiescent centre, and the cap initials are particularlysensitive. In Vicia the main change in mitosis is that the quiescentcentre increases its rate, but at 12°C there is a slightstimulation of division all over the meristem. In Vicia increasingthe dose rate or lowering the temperature increases the nucleardamage. At 19° C there is an increase in damage with accumulateddose percell cycle, but the data at 12°C do not fall onthe 19°C curve, suggesting that there may be a temperatureeffect on damage other than that caused by changing the durationof the cell cycle. The differences in radiosensitivity betweenthe different regions of the meristem are due solely to differencesin the rates of mitosis of the cells.     

Effect of abscisic acid on the cell cycle in the growing maize root

The mechanism by which the rate of cell proliferation is regulated in different regions of the root apical meristem is unknown. The cell populations comprising the root cap and meristem cycle at different rates, proliferation being particularly slow in the quiescent centre. In an attempt to detect the control points in the cell cycle of the root apical meristem of Zea mays L. (cv. LG 11), quiescent-centre cells were stimulated to synthesise DNA and to enter mitosis either by decapping or by immersing intact roots in an aqueous 3,3-dimethyl-glutaric acid buffer solution. From microdensitometric and flow-cytometric data, we conclude that, upon immersion, the G₂ phase of the cell cycle of intact roots was shortened. However, when 50 M abscisic acid (ABA) was added to the immersion buffer, parameters of the cell cycle were restored to those characteristic of intact roots held in a moist atmosphere. On the other hand, decapping of primary roots preferentially shortened the G₁ phase of the cell cycle in the quiescent centre. When supplied to decapped roots, ABA reversed this effect. Therefore, in our model, applied ABA retarded the completion of the cell cycle and acted upon the exit from either the G₁ or the G₂ phase. Immersion of roots in buffer alone seems to trigger cells to more rapid cycling and may do so by depleting the root of some ABA-like factor.Abbreviations ABA cis-abscisic acid - DGA 3,3-dimethyl-glutaric acid - DAPI 4,6-diamidino-2-phenylindole - LI labelling index We thank Pierre Zaech of the Ludwig Institute, Epalinges, Switzerland, for expert assistance in flow cytometry and Dr. Jean-Marcel Ribaut of our Institute for providing data on exodiffusion and metabolism of ABA.     

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     

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 extent and differences in mitotic activity of the root tip ofVicia faba L.

Mitotic activity does not stop for different meristematic cells of the root apex at the same distance from the initials. The differences are connected with the functional heterogeneity of the apical meristem of the root. The arrangement of vascular bundles,i.e. the alternation of independent xylem and phloem groups, is of major importance. In broad bean roots, the protophloem sieve elements stop dividing first. The centre of the stelei. e. late metaxylem elements stop dividing next. Division in the stele gradually ceases centrifugally, while it ceases centripetally in the peripheral part of the root. The cylindrical region with prolonged cell division includes internal layers of the cortex including endodermis, pericycle and adjoining cells of the stele. Proximally apical meristem is reduced to isolated strands of cells adjacent to the protoxylem poles. Pericycle cells stop dividing last at a distance of approx. 9–10 mm from the initials. The number of the division cycles is limited and is specific for individual cell types. Epidermal and cortical cells divide in broad bean roots transversely approximately seven times, cells of late metaxylem approximately five times. Root apical meristem is an asynchronous cell population with a different duration of the mitotic cycle. We determined local variations in the duration of the mitotic cycle in the apical meristem of broad bean root by means of colchicine-induced polyploidy. The cells of the quiescent centre had the longest mitotic cycle after colchicine treatment. The region of the proper root adjacent to the quiescent centre was mixoploid (2n and 4n). Isolated cells with a long cycle occurred also in the cortex and in the central cylinder. Cells with a division cycle of 18h were found in the root cap, in the epidermis, in the cortex and in the central cylinder. Relatively numerous cells with the shortest division cycle, approx. 12 h, occurred farther of the quiescent centre in the epidermis, in the cortex, in the pericycle, and in adjacent layers of the stele through-out the entire meristematic region. The results derived from the analysis of the apical meristem are discussed in connection with the ontogenesis of different types of cells taking part in the primary structure of the root.     

Meristem-Specific Suppression of Mitosis and a Global Switch in Gene Expression in the Root Cap of Pea by Endogenous Signals

Two functionally distinct sets of meristematic cells exist within root tips of pea (Pisum sativum): the root apical meristem, which gives rise to the body of the root; and the root cap meristem, which gives rise to cells that differentiate progressively through the cap and separate ultimately from its periphery as border cells. When a specific number of border cells has accumulated on the root cap periphery, mitosis within the root cap meristem, but not the apical meristem, is suppressed. When border cells are removed by immersion of the root tip in water, a transient induction of mitosis in the root cap meristem can be detected starting within 5 min. A corresponding switch in gene expression throughout the root cap occurs in parallel with the increase in mitosis, and new border cells begin to separate from the root cap periphery within 1 h. The induction of renewed border cell production is inhibited by incubating root tips in extracellular material released from border cells. The results are consistent with the hypothesis that operation of the root cap meristem and consequent turnover of the root cap is self-regulated by a signal from border cells.     

Localization of Nucleic Acid Synthesis in Root Meristems

Adenine-8-C1¹⁴ was supplied to roots of Vicia faba and Alliumascalonicum and its incorporation into DNA was studied fromautoradiographs of hydrolysed sections. These roots have a quiescentcentre to the meristem where the cells do not synthesize DNAand probably, therefore, play no part in the construction ofthe root. The boundary between the quiescent centre and thecentral cap initials is clearly denned and this suggests thatthere is as little cell interchange between the histogens asthere is in roots with visibly discrete histogens.     

Correlations Between the Dimensions of Different Zones of Grass Root Apices, and their Implications for Morphogenesis and Differentiation in Roots

The sizes of different zones within root apices of nine speciesof grass were estimated, and statistically significant correlationswere found between certain of them. The volume of the cap isrelated to the volume of the meristem of the root proper. Thecortical and stelar portions of the meristem are also related,and their lengths and volumes correlate with the volume of thequiescent centre. The volume of the quiescent centre also correlateswith the length of the zone in which periclinal divisions arefound in the inner cortex; these divisions generate the rowsof cells across the cortex. Diameter of the procambial cylinder,quiescent centre volume and vascular complexity are related,though from correlations alone it is not possible to say whetherone of these characters directly influences another as has beensuggested by other workers. All the zones within the root apexprobably form a tightly-integrated developmental unit. Root structure seems to be independent of cell size, thoughcell size correlates with nuclear DNA content which is a species-specificfeature. Gramineae, meristems, morphogenesis, root apices     

3D analysis of mitosis distribution highlights the longitudinal zonation and diarch symmetry in proliferation activity of the Arabidopsis thaliana root meristem

To date CYCB1;1 marker and cortex cell lengths have been conventionally used to determine the proliferation activity of the Arabidopsis root meristem. By creating a 3D map of mitosis distribution we showed that these markers overlooked that stele and endodermis save their potency to divide longer than the cortex and epidermis. Cessation of cell divisions is not a random process, so that mitotic activity within the endodermis and stele shows a diarch pattern. Mitotic activity of all root tissues peaked at the same distance from the quiescent center (QC); however, different tissues stopped dividing at different distances, with cells of the protophloem exiting the cell cycle first and the procambial cells being the last. The robust profile of mitotic activity in the root tip defines the longitudinal zonation in the meristem with the proliferation domain, where all cells are able to divide; and the transition domain, where the cell files cease to divide. 3D analysis of cytokinin deficient and cytokinin signaling mutants showed that their proliferation domain is similar to that of the wild type, but the transition domain is much longer. Our data suggest a strong inhibitory effect of cytokinin on anticlinal cell divisions in the stele.     

Extra DNA synthesis in the dedifferentiating cells ofVicia faba roots

Summary Cell dedifferentiation was induced inVicia faba root tissues by removing the whole root meristem (decapitation) and the behaviour of the nuclear DNA in the dedifferentiating cells was studied by means of cytophotometric and autoradiographic analyses. Cytophotometric determination after Feulgen-staining showed that: 1. the vast majority of nuclei in differentiated cells were in the DNA postsynthetic phase, but their Feulgen absorption was lower than that of DNA postsynthetic nuclei (G₂, 4 C) in the meristem; 2. such a Feulgen absorption was detected in certain nuclei after root decapitation; 3. all the mitoses in the dedifferentiating tissues were diploid, fully matching the Feulgen absorption of mitoses in the meristem.After³H-thymidine (³H-T) feeding of the decapitated roots and autoradiography, the following results were obtained: 1. two populations of labeled nuclei, characterized by two different levels of scattered labeling occurred in dedifferentiating tissues, slightly labeled nuclei being much more numerous than heavily labeled nuclei; 2. the percentage of labeled nuclei was much greater than that of DNA presynthetic nuclei in the root tissues; 3. almost all the mitoses were labeled after a 16-hour³H-T feeding; 4. the percentage of slightly labeled nuclei paralleled that of dedifferentiating cells; 5. the duration of the DNA synthesis phase and that of the gap between completion of DNA synthesis and mitosis differed in heavily and slightly labeled nuclei; 6. all nuclei which entered DNA synthesis also entered mitosis.These results are interpreted to mean that: 1. after decapitation, two different DNA syntheses occur in the dedifferentiating root tissues ofV. faba: DNA reduplication in cells which dedifferentiate starting from a DNA presynthetic nuclear condition (heavily labeled nuclei) and extra DNA synthesis in cells which dedifferentiate starting from a DNA postsynthetic nuclear condition (slightly labeled nuclei); 2. extra DNA synthesis is required in these dedifferentiating cells for entry into mitosis.     

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.