Experimental and Analytical Studies of Pteridophytes: XXXII. Further Investigations on the Effect of Undercutting Fern Leaf Primordia

An account is given of experiments in which young primordiaand primordium sites at the apex of Dryopteris aristata wereisolated basally and laterally by undercutting and deep radialincisions. The treatments had the effect of destroying or severingthe incipient vascular tissue underlying the primordium on itsabaxial and lateral sides. Provided the shoot apex had not beendamaged during treatment, neither buds nor centric (or radial)leaves were induced as a result of the experimental procedureemployed. On the contrary leaves with distended bases, showingan abnormally rapid rate of growth, were usually produced. Theresults obtained are against the view that a leaf-trace, differentiatingacropetally in advance, determines the formation of a primordium;but they support the thesis that the apical cell group exercisesa regulative effect on the growth and morphogenetic activitiesof the apical meristem and young primordia. The evidence alsosupports the view that the incipient vascular tissue is importantin morphogenesis in that it affords a pathway for the translocationof nutrients and hormonal substances.     

The Relationship Between the Distribution of Periclinal Cell Divisions in the Shoot Apex and Leaf Initiation

Periclinal cell divisions in vegetative shoot apices of Pisumand Silene were recorded from serial thin sections by mappingall the periclinal cell walls formed less than one cell cyclepreviously. The distribution of periclinal divisions in theapical domes corresponded to the distributions subsequentlyoccurring in the apices when the young leaf primordia were forming.In Pisum, periclinal divisions were almost entirely absent fromthe I₁ region of the apical dome for half a plastochron justafter the formation of a leaf primordium and appeared, simultaneouslyover the whole of the next potential leaf site, about half aplastochron before the primordium formed. In Silene periclinaldivisions seemed to always present in the apical dome at thepotential leaf sites and also round the sides of the dome wherethe ensheathing leaf bases were to form. Periclinal divisionstherefore anticipated the formation of leaf primordia by occuring,in Pisum about one cell cycle and in Silene two or more cellcycles, before the change in the direction of growth or deformationof the surface associated with primordial initiation. Pisum, Silene, planes of cell division, orientation of cell walls, leaf primordia, shoot apical meristem, plastochron     

Determination and Differentiation of Leaf and Petal Primordia in Impatiens balsamina

The development of primordia as leaves, petals, or as organsintermediate between leaves and petals can be regulated by photoperiodin Impatiens. In intermediate organs only some parts of theorgan differentiated as petal, and then only in some cell layers.Allometric measurements of primordium shape suggested that intermediateorgans may begin development as petals, and that their intermediatecharacter at maturity resulted from a switch of some parts ofthe organs from petal to leaf development when the primordiawere between 0.5 and 1 mm long. In reverted apices made to re-flower,primordia were not completely determined as leaves until theywere about 750 µm long. Determination typically occurredfirst at the tips and last at the bases of these primordia.The determination of primordia as leaves or petals in Impatiensis discussed in relation to primordium determination in otherspecies. It is suggested that the lack of commitment to flowermay result in relatively late primordium determination in Impatiens. Impatiens balsamina, determination, differentiation, leaf and petal development, flowering, reversion     

Effects of Developing Leaves on Stelar Pattern Development in the Shoot Apex of Matteuccia struthiopteris

A developmental study of the normal shoot apex of Matteucciastruthiopteris suggested that patterned stelar differentiationis initiated immediately beneath the single layer of promeristemand occurs prior to the initiation of the youngest leaf primordium.A developmental study in which all leaf primordia were suppressed,with or without lateral isolation of the terminal meristem byvertical incisions, has confirmed this interpretation of stelardifferentiation. Experimentally-induced changes in the tissueimmediately below the promeristem were reflected in the resultingmature structure of the stele. Failure of leaf gap initialsto differentiate, if all leaf primordia were suppressed at theincipient stage, resulted in a mature stele without leaf gaps.Similarly the disappearance of pith mother cells after severalweeks of leaf removal was associated with the formation of astele without pith. Leaf influence was further assessed by allowingone primordium to develop while all others were suppressed.The developing leaf had a small promoting effect on caulinevascular tissue differentiation but its major impact on theexpansion of the parenchymatous tissues of the stele. Characteristicprotoxylem and protophloem failed to differentiate when allleaves were suppressed and, when leaf was allowed to develop,formed only in relation to the leaf.Copyright 1995, 1999 AcademicPress Leaf influence, vascular pattern formation, experimental surgery, shoot apex development, protoxylem, protophloem, Matteuccia struthiopteris     

Following the initiation and development of individual leaf primordia at the level of the shoot apical meristem: the case of distichous phyllotaxis in Begonia

BACKGROUND AND AIMS: By using the technique of replicas of a developing apex it is possible to obtain a direct measure of phyllotactic parameters (plastochrone and platochronic ratio) involved in the initiation of two successive primordia at the level of the SAM. The goal of this study is to compare, in a real time setting, the value of phyllotactic parameters in distichous systems using Begonia as a case study, with the value of the same parameters in spiral phyllotactic systems. METHODS: To determine the real-time sequence of events at the level of the SAM, replicas were made of the developing apex at different intervals using previously described techniques. Impression moulds were made at 24-h intervals. The following phyllotactic parameters were measured: plastochrone, angle of divergence, plastochrone ratio and ratio between the diameter of the leaf and the apex. RESULTS: The time between the appearance of two successive leaves is 15-20 d. The average value of the plastochrone ratio (R) is 1.3, and the ratio of the leaf to the diameter of the apex (Gamma) is 2.5. The angle of divergence varies from 165 masculine to 180 masculine. The speed of advection of the primordium from the apex, varies from 0.28 to 0.37 microm d(-1). CONCLUSIONS: The speed of advection of primordia in Begonia is lower than that of Anagalis. This is not in accordance with theoretical simulations that predict the opposite. In Begonia, the plastochrone ratio does not reflect the real time of appearance of two successive primordia. The time separating the appearance of two primordia is not directly related to the distance of these two primordia from the centre of the apex but is related instead to the enlargement of leaves.     

Experimental and Analytical Studies of Pteridophytes: XXVIII. Leaf Symmetry and Orientation in Ferns

An account is given of surgical treatments which may determinethe symmetry and orientation of leaf primordia in ferns. Inone series small deep tangential incisions, severing the incipientvascular tissue, were made immediately above very young leafprimordia or primordium sites: the primordia developed as leavesin all cases. In the other series two deep and wide tangentialincisions were made above a primordium (or site) so as to leavea ‘bridge’ of intact tissue between it and the apicalcell of the shoot. In some of the more actively growing apicesbuds were formed, even though the shoot apical cell was quiteundamaged; relatively inactive apices typically yielded leaves.It is concluded that the orientation and symmetry of a leafprimordium cannot be referred only to the direct action on itof a growth-regulating substance moving basipetally from theapical cell, but rather that these characteristic developmentsare mediated through the organization and physiological activityof the apex as a whole, the intact apical cell being a centraland essential element of the system.     

Identification of Shoot Apical Meristem Cells Committed to Form Vascular Elements in Pisum sativum L. and Vicia faba L.

A cytochemical study of naphthol AS-D esterases in vegetativeshoot apices of Pisum sativum and Vicia faba L. has shown thepresence of carboxyl esterases (E.C. 3.1.1.1 [EC] .) in those meristemcells already committed to form vascular elements. These cellsform a sequence linking the morphologically identifiable procambiumto the cells of the tunica layers at a site either already identifiableas the next primordium or which will form the next primordium.The implications of this result are briefly discussed in relationto the control of primordia formation and procambial cell development. Pisum sativum, Vicia faba, determination, vascular tissue, shoot apex, cytochemistry     

Rates of Leaf Initiation and Leaf Growth in Agropyron repens (L.) Beauv.

Rates of leaf initiation, emergence, and growth have been measuredduring the period between the production of one and of ten matureleaves on the primary shoot of Agropyron repens. There is aprogressive decline in the growth rate of successively formedleaves so that at the time of formation of the next leaf primordiumeach primordium is smaller than its predecessor at a comparablestage of development. There is also a trend towards a diauxicpattern of growth with a lag phase apparently coinciding withthe transition from apical to intercalary growth of the youngleaf. Up to the six-leaf stage the rate of leaf formation exceedsthe rate of emergence and leaf primordia accumulate on the shootapex. Thereafter the rate of emergence exceeds the rate of formation.These changes in rates of leaf formation and growth are interpretedin terms of competition for assimilates between expanding leavesand leaf primordia, and between the primary and axillary shootapices.     

Initial Differentiation of Vascular Tissue in the Shoot Apex of Carrot (Daucus carotaL.)

Although an initial stage of vascular differentiation precedingprocambium has been demonstrated in ferns, its presence in seedplants has not been accepted generally. In the shoot apex ofcarrot, a short cylinder of provascular tissue is recognizedas the initial stage of vascular differentiation. This firstbecomes apparent through the enlargement and vacuolation ofpith and cortical tissue rather than as a result of specificchanges in the provascular tissue itself. Procambium in discretestrands differentiates acropetally in the provascular tissuein relation to developing leaf primordia. Provascular tissueis not recognized above the axil of the youngest leaf primordiumbut it is distinct at or above the level at which the traceof the youngest primordium diverges. Support for the recognitionof provascular tissue is provided by a positive reaction tohistochemical tests for carboxylesterases in this tissue aswell as in procambium and later stages of vascular differentiation.Copyright1999 Annals of Botany Company. Provascular tissue, carboxylesterase, shoot apex, vascular differentiation, carrot (Daucus carotaL.).     

The Effect of Temperature on the Initiation of Leaf Primordia in Developing Potato Sprouts

Kirk, W W., Davies, H. V. and Marshall, B. 1985. The effectof temperature on the initiation of leaf primordia in developingpotato sprouts.—J. exp. Bot. 36: 1634–1643. Initiation of leaf primordia in potato sprouted out of soilin light was an asymptotic function of thermal time and thebase temperature for the process was 3.6 °C. The parametervalues of the asymptotic function were universal for cv. MarisPiper. The estimated rate of leaf primordium initiation decreasedlinearly from 0.033 leaf pnmordia (K day)^–1 when abouteight leaf primordia were present to zero after a maximum numberof 24 leaf primordia had been initiated. The decrease in rateof development with increasing number of primordia may be dueto depletion of mother tuber resources. The transition of theapex from a vegetative to a reproductive state was not the factorlimiting the initiation of additional leaf primordia. Key words: Potato, Solanum tuberosum L., leaf primordia initiation, temperature, thermal time, development     

Surgical Experiments on the Differentiation of Vascular Tissue in the Shoot Apex of Carrot (Daucus carota L.)

Surgical techniques were applied to the shoot apex of carrot(Daucus carota L.) to test the interpretation that provasculartissue is the initial stage of vascular differentiation andto localize the sources of the influences that control its differentiation.If the apex is isolated laterally by vertical incisions leavingit at the summit of a plug of pith tissue, vascular differentiationproceeds normally and an independent vascular system is formedin the pith plug. If all leaf primordia are systematically suppressed,provascular tissue continues to differentiate as an acropetalextension of the pre-existing vascular system but no furtherdifferentiation occurs. When the apex is isolated laterallyand all leaf primordia are suppressed, provascular tissue continuesto be formed acropetally and is extended basipetally into thepith plug by redifferentiation of pith cells, but no furtherdifferentiation occurs. This tissue reacts positively to histochemicaltests for esterase indicating its vascular nature. If only oneleaf primordium is allowed to develop on an isolated shoot apex,its vascular system develops normally and extends basipetallyinto the pith plug, but there is no extension of provasculartissue into the pith plug. These results support the interpretationthat the initial stage of vascular differentiation is controlledby the apical meristem but that further maturation of vasculartissue depends upon influences from developing leaf primordia.Copyright 2000 Annals of Botany Company Provascular tissue, differentiation, carrot (Daucus carota L.), shoot apex, surgical techniques, leaf primordia     

Leaf Ontogeny in Digitaria eriantha

The shoot apex consists of two layers, the dermatogen and thehypodermis. The leaf primordia arise through periclinal divisionswithin these two layers on the side of the apex. Further divisionsof the dermatogen push the little protuberance upward and togetherwith divisions the hypodermis add internal tissues of the youngleaf. When the median and lateral bundles of the primordia arisein Digitaria eriantha they are isolated from the vascular supplyof the rest of the plant. The median strand, the first to form,and the first order laterals form at the disc of insertion ofthe primordium. The other laterals form higher up in the primordium.These strands extend both acropetally and basipetally to linkwith the vascular supply of the rest of the plant. Digitaria eriantha, apical meristem, leaf primordium, vascular bundle, orange G, tannic acid, iron alum     

Chimeras and the Origin of Lateral Root Primordia in Zea mays

The difficulty of determining the contribution made by the pericycleand endodermis of mother roots to lateral primordia in the Gramineaehas been solved by inducing polyploid chimeras at initiation. The endodermis forms a layer covering the primordium, but thislayer does not form the epidermis of the lateral. It does formthe root cap of the young primordium, but this is replaced ata variable stage of development by the quiescent centre donatinga new set of cap initials of pericyclic origin. Reasons forthe previous diversity of interpretations are presented. chimera, lateral root primordia, Zea mays     

Cortical Cell Breakdown and Lateral Root Primordium Development in Vicia faba L

The effect of the formation of a cavity in the cortex of theprimary root of Vicia faba adjacent to lateral root primordiaon root development has been investigated. Premature exposureof such primordia to the external medium by removing the overlyingtissues of the primary root has no effect on primordium developmentif that primordium was within 48 h of emerging as a lateralroot. Similar exposure of primordia which were at an earlierstage of development and consisted of between 3400 and 7000cells resulted in the generation of a stationary phase, withmost of the nuclei arrested in G1 (presynthetic interphase),48–72 h after exposure began, followed by nuclear degenerationby 96 h. Since no mature vascular tissue was found in theseprimordia until after they emerged as secondary roots, all ofthe nutrients necessary for the maintenance of cell proliferationin these meristems must reach them by simple diffusion fromthe surrounding medium. A preliminary analysis of the liquidcontents of the cavity next to developing primordia demonstratesit to be rich in carbohydrates and it is clear, from the resultsreported in this paper, that cell proliferation in primordia,consisting of a mean number of 5400 cells, is largely dependenton the substances present in the cavity fluid, although somematerials reach the primordium by diffusion from the cells ofthe primary root to which the primordium remains attached.     

Independence of Organogenesis and Cell Pattern in Developing Angle Shoots of Selaginella martensii

Angle meristems are mounds of meristematic tissue located atdorsal and/or ventral branch points of the dichotomising stemaxes of many species of Selaginella (Lycophyta). The presentstudy examined the development of ventral angle shoots of S.martensii in response to removal of distal shoot apices (decapitation).Scanning electron microscopy of sequential replicas of developingangle meristems and angle shoots revealed that for the firsttwo pseudowhorls of leaf primordia, particular leaves are notattributable to particular merophytes of the angle meristemapical cell. Individual leaf primordia of the first (outer)pseudowhorl often form from more than one merophyte. Neitherthe shape of the angle meristem apical cell nor the directionof segmentation has any effect on the development of the angleshoot. Additionally, the apical cell of the angle meristem doesnot necessarily contribute directly to either of the new shootapices of the developing angle shoot. The first bifurcationof the angle shoot shows a remarkably consistent relationshipto the branching pattern of the parent shoot. The strong branchof the first angle shoot bifurcation typically occurs towardthe weak side branch of the parent shoot. Anatomical studiesshowed that bifurcation of the young angle shoot involved theformation of two new growth centres some distance away fromthe original angle meristem apical cell; new apical cells subsequentlyformed within these. These results provide additional supportfor the view that cell lineage has little or no effect on finalform or structure in plants.Copyright 1994, 1999 Academic Press Selaginella martensii Spring, Lycophyta, angle meristem, apical cell, shoot apical meristem, leaf primordium, branching, dichotomy, morphogenesis, determination, competence, development, mould and cast technique, replica technique, scanning electron microscopy     

The Shoot Apex and its Dichotomous Branching in the Nypa Palm

Seedlings of the palm Nypa fruticans van Wurmb are viviparous,the plumule becoming exserted before the fruit is ripe and possiblyassisting in fruit detachment. Established seedlings have horizontalaxes, this growth orientation being maintained throughout thelife of the palm which may be described as ‘rhizomatous’.Inflorescences are axillary in adult plants, but their distributionis irregular. The shoot apex is small and very asymmetricalsince it is more or less displaced into a lateral position byprogressive enlargement of each leaf primordium during thisleaf's first plastochrone. The plastochrone interval is apparentlya long one so that leaves have a wide developmental gap betweenthem. This results in a leaf base which is more or less cylindricalbut with a groove to accommodate the next youngest leaf. All the available evidence suggests that vegetative branchingis by equal dichotomy of the shoot apex at wide intervals. Thedichotomy is marked superficially by a leaf, enclosing the twonew shoots, which has two grooves. The twin shoots are insertedin the lateral, not the dorsiventral plane of this enclosingleaf. The daughter shoots are always at precisely the same stageof development and they always have mirror-image symmetry withrelation to each other, the phyllotaxis of the parent shootbeing maintained without obvious interruption. There is no anatomicalevidence for abortion of the original apex and its replacementby two new ones. The vascular system is divided equally betweeneach daughter shoot without interruption, suggesting stronglythat there is continuity of growth from the undivided to thedivided condition.     

Initiation of the Vascular System and the Transition to Flowering in Early Tassels of Zea mays Land Race Chapalote (Poaceae)

Serial transections of young tassels of (Zea mays land race) chapalote revealed relationships between the vascular system in its procambial state and the lateral primordia along the axis. A lateral tassel primordium usually consists of an indefinite rim with a prolongation that will become a tassel branch or spikelet pair. A lateral tassel primordium usually develops via modifications of the vegetative leaf primordium in which the leaf apex is enhanced but the leaf base and the bud it produces are suppressed. The clearest sign of the transition from the vegetative state to the tassel is the scale leaf, which is intermediate in form between a vegetative leaf and a lateral tassel primordium. Procambial traces differentiate in isolation in the tassel axis in response to the lateral tassel primordia. Adjacent procambial traces then link axially into sympodia to initiate the three-dimensional vascular system of the tassel axis. Older sympodia occur near the center of the axis interior to more recently initiated procambial traces. Procambial continuity does not occur between the tassel axis and the lateral primordia until isolated traces in the lateral primordia link with the sympodia in the tassel axis. The transition from distichy to polystichy by the lateral tassel primordia occurs as the narrowing of the leaf base makes space available on the tassel axis for lateral primordia out of the vegetative distichous plane.     

A Sequential Study of Cell Divisions and Expansion Patterns on a Single Developing Shoot Apex of Vinca major

During the growth of a single developing vegetative apex ofVinca major, both the orientation and frequency of cell divisions,and the pattern of cell expansion, were observed using a non-destructivereplica technique. Micrographs taken at daily intervals illustratethat the central region of the apical dome remains relativelyinactive, except for a phase of cell division which occurs after2 d of growth. The majority of growth takes place at the proximalregions of the dome from which develop the successive pairsof leaves. The developing leaf primordia are initiated by aseries of divisions which occur at the periphery of the centraldome and are oriented parallel to the axis of the subsequentleaves. The cells which develop into the outer leaf surfaceof the new leaves undergo expansion and these cells divide allowingfor the formation of the new leaf. This paper describes thefirst high-resolution sequential study of cell patterns in asingle developing plant apex. Sequential development, cell division, expansion patterns, SEM, Vinca major, apical dome, leaf primordium, leaf initiation     

Experimental and Analytical Studies of Pteridophytes: XXXV. The Effects of Direct Aplications of Varous Substances to the Shoot Apex of Dryopteris austriaca (D. aristata)

The organization and morphogenetic activity of a fern apex,that of Dryopteris austriaca (D. aristata), have been furtherinvestigated by observing how it responds to direct applicationsof solutions of various physiologically active organic substances.Large apices were laid bare and treated for periods of up toabout 20 days with solutions of 3-indoleacetonitrile, dinitrophenol,maleic hydrazide, yeast extract, glutamine, &c., with directexamination at frequent intervals using a binocular microscope. In most cases some reaction was obtained. Developments of morphogeneticinterest include: an increase in the rate of growth in the subapicalregion; an arrest or slowing down of growth in the apical meristem;the formation of flattened and of sunken, cup-shaped apices;the formation of double, treble, and quadruple leaf primordia,of two primordia in a single leaf site, and of centric (or radial)leaves; the occasional suppression of primordia; the reversalof the phyllotactic spiral; the induction of buds on meristemswith an uninjured apical cell; the promotion of root formationand growth; the formation of scales on the apices of young leafprimordia and leaf sites, and the precocious formation of scalesover the whole of the apical meristem, including the apicalcell; and the precocious parenchymatization of the meristemincluding the apical cell. The significance of these resultsis discussed.     

Model for the role of auxin polar transport in patterning of the leaf adaxial–abaxial axis

Leaf adaxial–abaxial polarity refers to the two leaf faces, which have different types of cells performing distinct biological functions. In 1951, Ian Sussex reported that when an incipient leaf primordium was surgically isolated by an incision across the vegetative shoot apical meristem (SAM), a radialized structure without an adaxial domain would form. This led to the proposal that a signal, now called the Sussex signal, is transported from the SAM to emerging primordia to direct leaf adaxial–abaxial patterning. It was recently proposed that instead of the Sussex signal, polar transport of the plant hormone auxin is critical in leaf polarity formation. However, how auxin polar transport functions in the process is unknown. Through live imaging, we established a profile of auxin polar transport in and around young leaf primordia. Here we show that auxin polar transport in lateral regions of an incipient primordium forms auxin convergence points. We demonstrated that blocking auxin polar transport in the lateral regions of the incipient primordium by incisions abolished the auxin convergence points and caused abaxialized leaves to form. The lateral incisions also blocked the formation of leaf middle domain and margins and disrupted expression of the middle domain/margin‐associated marker gene WUSCHEL‐RELATED HOMEOBOX 1 (SlWOX1). Based on these results we propose that the auxin convergence points are required for the formation of leaf middle domain and margins, and the functional middle domain and margins ensure leaf adaxial–abaxial polarity. How middle domain and margins function in the process is discussed.