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     

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.     

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     

Organelle DNA Synthesis in the Quiescent centre of Arabidopsis thaliana (Col.)

The behaviour of cell nuclei and organelle nucleoids (organellenuclei) was studied in the root apical meristem of 3-d-old seedlingsof Arabidopsis thaliana (Col.). Samples were embedded in Technovit7100 resin, cut into thin sections and stained with 4'-6-diamidino-2-phenylindole(DAPI) for observation of DNA. DNA synthesis in cell nucleiand organelle nucleoids was investigated using the incorporationof [³H] thymidine or 5-bromo-2'-deoxyuridine (BrdU). Incorporated[³H] thymidine and BrdU were detected by microautoradiographyor immunofiuorescence microscopy, respectively. Central cellsand cells just above the central cells of the quiescent centre(QC) showed an extremely low activity of DNA synthesis. However,DNA synthesis occurred in at least one organelle nucleoid ofall cells in the QC within 24 h. This suggests the cells inthe QC are quiescent with regard to nuclear DNA synthesis, butnot with regard to the organelle nucleoids. Key words: Arabidopsis thaliana, quiescent centre, root apical meristem, mitochondrial nucleoid (nuclei), plastid nucleoid (nuclei)     

Determination of Stelar Elements in Roots of Pisum sativum L.

Cytochemical studies of esterase activity in 0.5 mm segmentsfrom root tips of Pisum sativum explanted for up to 9 days inbasal culture medium containing 2 per cent sucrose showed retentionof this activity. During this time, all segments from the secondand third 0.5 mm segments of the root tip developed xylem elementsas did the proximal end of the first segment. No xylem elementswere found in the 12–14 cells behind the quiescent centre.It is concluded that the central group of meristem cells aregenerally programmed to form tissues of the stele immediatelyon leaving the quiescent centre, and that the programming forxylem and phloem elements occurs as a second step. Pisum sativum L., garden pea, determination, histochemistry, esterases, stele, root     

Micronuclei and Radiosensitiviy in the Root Meristem of Vicia faba

Percentages of cells with micronuclei in four regions of theroot meristems of Vicia faba are used as measures of sensitivityto acute X-irradiation. There are two peaks in these percentages,occurring at about four and eight days after 360 rads and twoand six days after 180 rads. Two peaks exist, probably becausethe radiation delays cells that were in G₁ much more than cellsthat were in G₂ in reaching the first post-irradiation mitosisand consequently in displaying micronuclei in the followinginterphase. The relative heights of the two peaks thereforereflect the relative numbers of cells in G₁ and G₂ as well asthe relative sensitivity of the two phases to chromosomal damage. The cells of the quiescent centre are injured least by the radiationas they are mostly held at Gt. The meristem thus obeys the lawof Bergonié and Tribondeau, but differs from that ofZea in that the meristematic cells of the cap initials and steleimmediately adjacent to the quiescent centre resemble the quiescentcentre much more closely than the stele 250 µ away inthe numbers of micronuclei produced. This is consistent withthe differences already known between the two species concerningrates of division in the different regions of the meristem andthe behaviour of the meristem after severe radiation injury.     

Proliferating and Quiescent Cells in the Apical Meristem of Elongating Lateral Roots of Vicia faba L.

Whole root systems of Vicla faba were continuously exposed to³H-TdR for periods of up to 72h, following which LI was determinedin the cap initials, in the quiescent centre or in that partof the meristem in which a quiescent centre will develop, andin the stele and in the cortex-epidermis at intervals alongthe apical 800 µm basal to the junction between the capinitials and the rest of the meristem, in newly-emerged (NE),0.2 and 4.0 cm long lateral roots, after each exposure period.Cell doubling time (Td), mean cycling time (Tc) and the sizeof the growth fraction (GF) were then calculated for each partof the meristem investigated in each batch of roots, from thecurves recording increase in labelling index (LI) with increasein duration of the period of exposure to ³H-TdR and from therate of increase in LI over the initial l-12h labelling period.Since it is extremely difficult to eliminate all sources oferror in calculating GF from the values obtained for LI in continuouslabelling experiments, it is emphasized that the values of GFreported in the present paper may not be totally accurate. Thisis also true of the results obtained for Tc as Tc was derivedfrom the product of the corresponding values for Td and GF. Cell doubling time and mean cycling time were both longer inthe cells forming the quiescent centre in the 0.2 and 40 cmlong roots than in any other part of the apical meristem examined.The size of the GF was found to decrease basally along the steleand the cortex-epidermis from the most apical to the most basalsegment examined in the NE, while Td increased in duration.Similar changes took place along the stele of the 0.2 cm longlaterals, but not in the cortex-epidermis of these roots or,to any great extent, in any of the tissues examined in the 4.0cm secondary roots. No consistent trend was apparent in theduration of Tc basally along either tissue examined in theseroots. It was concluded from these results, and from supportingdata in the literature, that, as the laterals elongated fromNE to 4.0 cm, the apical meristem increased in length.     

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.     

The Meristem and Quiescent Centre in Cultured Root Apices of the gib-l Mutant of Tomato (Lycopersicon esculentum Mill.)

Cultured root apices of tomato bearing the gib-I mutation, whichreduces the levels of endogenous gibberellins, grew slower andwere thicker than wild-type contols. This was the result ofshorter and broader cells in the menstem of the mutant. Cellsof both cortex and stele were affected, but this did not causeany alteration to the volume fraction occupied by these twotissues in the root meristem. Root caps were longer in the mutantand there were also more layers of rhizodermis. All these effectscould be reproduced in wild-type roots by addition of 0.1µM2S, 3S paclobutrazol (an inhibitor of gibberellin biosynthesis)to the culture medium and could be normalized in mutant rootsby 0.1 µM GA₃. Cell doubling times in the proximal regionof the meristem were similar in mutant and wild-type roots,but were faster in both the quiescent centre (QC) and the capmeristem of the mutant. This latter feature of the mutant rootsis likely to be the cause of their longer caps, while the fasterrate of division in the QC accounts for the additional tiersof cells that were found to build up in the cortical portionof this zone These additional tiers failed to form in mutantroots grown in GA₃, but they could be induced in wild-type rootsby 2S, 3S paclobutrazol. These results suggest that endogenousgibberellins may be partly responsible for the slow rate ofcell growth and proliferation in the QC. Gibberellins, gib-I mutation, Lycopersicon esculentum, meristem, roots, 2S, 3S paclobutrazol, quiescent centre, tomato     

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.     

Protein Synthesis in Root Meristems

Comparative rates of incorporation of sulphate and L-leucmeinto proteins have been determined from autoradiographs forthe cells of root apices. This has confirmed the existence inmost roots of a quiescent centre where the rate of synthesisis much lower than in the surrounding cells. A more detailedaccount of leucine incorporation in the roots of Zea has beenmade by counting silver grains in quadrats on the autoradiographs.The rate of incorporation per unit volume increases on eitherside of the quiescent centre and then falls. On the proximalside the lowest rate of incorporation is below that in the quiescentcentre and is reached at 1–2 mm. from the tip. Fartherfrom the tip the rate rises. The rate of incorporation per celldoes not alter much in the first a mm. of the meristem, butbeyond that the rate rises steeply to between 15- and 5o-foldat 3 mm. from the tip. The patterns of change in rates of incorporationare different for the different tissues of the root.     

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.     

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 Fine Structure of the Quiescent Centre and Neighbouring Tissues3

The quiescent centre of the root meristem of Zea provides asource of undiffer-entiated, non-meristematic cells whose finestructure we have compared with that of the meristematic, non-differentiatedcells of the meristem and that of the differentiated, non-meristematiccells of the cap. We have shown an association between the developmentof the endoplasmic reticulum and the development of cell walls.Differentiation in the cap is accompanied by a decrease in thenumber of mitochondria and plastics per unit volume of cytoplasmand an increase in their number per cell. There is an even greaterincrease in the number of Golgi bodies per cell. The structureof the mitochondria becomes more elaborate in differentiatingcells especially in the cap and the Golgi increase their sizein meristematic and differentiating cells. These differencesare discussed in relation to some of the views about radiosensitivityand organization.     

Duration of the Mitotic Cycle in a Meristem

The length of the mitotic cycle and the time spent in mitosisare calculated for six regions of the apical meristem of rootsof Zea. The two methods used are the measurement of the ratesof labelling nuclei with radioactive thymidine and the ratesof accumulation of metaphases in roots grown in colchicine.The mitotic cycle lasts from 12 hours in the cap initials toabout 200 hours in the quiescent centre of the same roots. Somepoints about the organization of apices are discussed in thelight of these estimates.     

Origin of Epidermis and Development of Root Primordia in Pistia, Hydrocharis and Eichhornia

All three floating plants have roots bearing laterals derivedfrom both pericycle and endodermis. In Pistia and Eichhornialaterals arise within the meristem of the mother root; in Hydrocharisthey arise from mature tissue. In Pistia and Hydrocharis theepidermis becomes anatomically discrete between cortex and cap:in Pistia it is derived from the endodermis of the mother root,in Hydrocharis from the pericycle. The epidermis is not discretein Eichhornia and is derived from the pericycle of the motherroot with the cortex. Stathmokinetic data were used to construct timetables of developmentwhich show how the differences arise. In Pistia the first periclinaldivision of the endodermis-derived tissue individualizes theepidermis and occurs early, before a quiescent centre forms.In Hydrocharis the epidermis also becomes discrete before thepole of the meristem becomes quiescent, but it does so by apericlinal division of the pericycle-derived tissue. In Eichhorniapericlinal divisions occur in the outermost layer of the pericycle-derivedtissue long after quiescence has set in at the pole and afterthe fourth periclinal division in the endodermis derived cap.Its epidermis therefore never becomes anatomically discretethough it becomes functionally discrete because its polar cellsstop dividing as in the other plants. The involvement of the endodermis of mother roots in the formationof laterals is discussed in relation to the state of differentiationat sites of primordium formation, discreteness of the epidermisand subsequent fate of primordia. Pistia stratiotes L., Hydrocharis morsus-ranae L., Eichhornia crassipes Solms., primordia, lateral root, discrete epidermis, development, chimera, stathmokinetics     

Regeneration of the Root Cap of Zea mays L. and Pisum sativum L.: A Study with the Scanning Electron Microscope

Removal of the root cap from a root apex initiates regenerationof a new cap. The process has been followed using scanning electronmicroscopy. Quantitative data have been obtained for the growthin area of the exposed acroscopic surface of the quiescent centre(QC) and the increase in volume of the regenerating cap tissue.In Zea the surface of the QC shows an initial rapid increasein area followed by a slower increase. In Pisum the surfacearea increases uniformly, a rapid initial phase being absent.Together with observations on the behaviour of an incision atthe exposed surface, the results indicate that in Zea the capnormally imposes a constraint upon radial growth at the acroscopicsurface of the QC; in Pisum the QC appears not to be so constrained.The different responses may be related to the different arrangementsof cells at the apex of the meristem of these two species. Zea mays, Pisum sativum, maize, peao, scanning electron microscopy, root apex, regeneration     

Nuclear Chromatin Structure in Relation to Cell Differentiation and Cell Activation in the Cap and Quiescent Centre of Zea mays L.

Barlow, P. W. 1985. Nuclear chromatin structure in relationto cell differentiation and cell activation in the cap and quiescentcentre of Zea mays L —J. exp. Bot. 36: 1492–1503.Nuclear chromatin structure has been analysed by electron microscopyof thin sections of cells in four zones of the root cap—meristem,central, slime-secreting and outermost cells—and alsoin the quiescent centre of the root before and after decapping.The chromatin pattern has been related to the DNA and RNA syntheticactivity of the nuclei. During cap cell maturation there wasa progressive condensation of the chromatin and this was accompaniedby some reduction of RNA synthesis. The degree of condensationwas estimated from the area and number of pieces of electrondense chromatin which increased and decreased, respectively,during cap maturation. The volume fraction of condensed chromatinwas also estimated but, in the cap, was not found to be a goodindicator of nuclear activity. The outermost cells of the capshowed the greatest degree of chromatin condensation but werestill active in RNA synthesis. Microdensitometry of their nuclearDNA contents gave an indication of loss of DNA in some of thenuclei. Decapping activated DNA and RNA synthesis in the quiescentcentre and also stimulated a decondensation of chromatin: thenumber of condensed pieces of chromatin increased, and theirsize and volume fraction both decreased 4 h after decapping.The number of pores per unit length of nuclear envelope profilewas also estimated. In the cap this number increased duringcap maturation; in the activated quiescent centre the numberremained constant except for a small rise 4 h after decapping Key words: Zea mays, chromatin, root cap, quiescent centre     

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).     

The Quiescent Centre and Cell Determination in Roots of Pisum sativum

A combined autoradiographic and enzyme cytochemical study ofroot apices from Pisum sativum has permitted the demonstrationof a high naphthol AS-D esterase activity in the central groupof meristematic cells immediately outside the quiescent centre.This high naphthol AS-D esterase activity indicates an expressionof the programming of these cells to form elements of the stele.A consideration is given to the need for a quantal mitosis andof its possible occurrence in the quiescent centre. It is concludedthat the programming and mitotic activity may be two physiologicalevents which whilst occurring in the same cell, are not necessarilylinked events. Pisum sativum, roots, determination, quiescent centre, autoradiography, carboxylesterases