Transfusion Tissue in the Leaves of Thuja orientalis L.

The studies on the development and structure of transfusiontissue in the leaves of Thuja orientalis L. reveal that thetransfusion tissue develops all around the vascular bundle andforms a broad wing-like structure on either side of the midrib,in almost the entire length of the leaf. It extends beyond thevascular strand and reaches the apex where it is reduced considerably. The transfusion tissue in T. orientalis is of complex natureand is composed of living and non-living elements. The formerincludes transfusion parenchyma and albuminous cells while thelatter only the tracheids. Both vary in their size and shapeat various positions within a leaf.     

Effect of Excision on Leaf Water Potential

Improvements of thermocouple hygrometric techniques for in situleaf water potential measurement in the field now allows forcontinuous monitoring of water potential in response to an externalperturbation, such as leaf excision. Using Citrus jambhiri plants,measured leaf water potentials of completely excised leaf portionsimmediately increased when the petiole was excised or incisionswere made either transverse or parallel to the midrib. Incisionsparallel to the midrib were on the side nearest the hygrometeror opposite it if preceded by a petiole excision. Midrib incisionswere 100–150 mm long with the nearest cut edge being 20–50mm from the hygrometer cavity. All excisions were such thatleaf tissue was removed from the leaf with water potential onone of the leaf portions being measured continuously prior toand after excising. The peak increase in measured water potentialof the excised leaf portions ranged between 20 kPa and 80 kPabut averaged 50 kPa. In uncovered leaves, particularly underfield conditions with the associated high evaporative demand,measured leaf water potential declined rapidly after the initialincrease. The increase in measured water potential immediatelyfollowing various types of excision was confirmed for dark andlight conditions (laboratory and field respectively) using bothpsychrometric and dewpoint modes and occurred for secondaryexcisions, but to a lesser extent. Discovery of this phenomenonimplies that water potential measured on detached leaves maynot always represent accurately in situ leaf water potential. Key words: Leaf water potential, Thermocouple hygrometers, Leaf excision effects     

Ochna pulchra Hook: Leaf Growth and Development Related to Photosynthetic Activity

Ochna pulchra Hook. is a deciduous broad-leaved tree in theMixed Bushveld vegetation of the Northern Transvaal. The growthand development of leaves taken from trees in the field werestudied from a stage shortly before bud break, in late spring,until they were fully expanded and at the peak of photosyntheticactivity. Leaf area was measured by photographing the leaf against a transparentmm² grid. Finally a constant relationship between leaf area(A) and the linear dimension of length (L) and breadth (B) wasestablished: A = b x LB, where coefficient b = 0.72. Transverse sections of the lamina of the youngest leaves showeda five-layered plate meristem with a few functional conductingelements in the midrib. During further leaf development, celldivision was followed by means of autoradiography using [³H]thymidine.It was most active during the week after bud break. Leaf cell increment following on cell division made the majorcontribution to leaf growth resulting in a lamina that was atleast 90% expanded 4 weeks after bud break. The histologicalchanges accompanying cell division were observed using lightand electron microscopy. Even in late stages of leaf development mature and differentiatingstomata occurred together, limited to the abaxial epidermisand the midrib. Scanning electron microscopy showed stomataldistribution, their increasing density and gradual opening.The structure of these sunken stomata could reduce the outwarddiffusion of water vapour and increase the diffusion resistanceto carbon dioxide. Carbon assimilation rates of the developing leaves were measuredusing an IRGA (infra-red gas analyser) and their chlorophyllvalues were calculated. Photosynthesis was first measured amonth after bud break when the leaves were fully expanded, over50 % of the stomata exposed and leaf mesophyll tissue differentiatedwith mature chloroplasts. Net photosynthetic rates and chlorophyllvalues peaked 1 month later. Ochna pulchra Hook., photosynthesis, leaf development, leaf area, stomata, chlorophyll, savanna     

High ice nucleating ability in plant leaves

A high ice nucleating ability in leaves of Veronica persicaand Buxus microphylla was observed on analyzing the frequencydistribution of their freezing temperatures. The most efficientice nucleators in Veronica leaves exist in structures withinthe leaf blades (mesophyll), and those of Buxus in structureswithin the midrib (vascular tissue). Dissolved or suspendedleaf structures were much less effective. (Received March 16, 1973; )     

Leaf Heteroblasty in Pseudopanax crassifolius: Functional Significance of Leaf Morphology and Anatomy

Pseudopanax crassifolius (A. Cunn.) C. Koch. is a strongly heteroblastictree indigenous to New Zealand. Changes in the morphology andanatomy of leaves are described for seedling, juvenile, transitionaland adult phases of development. Seedlings produce five leaftypes; all are relatively small, thin, anthocyanic, have a lowspecific weight, and are anatomically comparable to the leavesof many shade plants. Juvenile leaves are long, linear, deflexed,coriaceous and sharply-toothed. They have a high specific weight,a thick, ornamented cuticle, a multiseriate hypodermis composedof collenchyma, well developed palisade and many spongy mesophylllayers. Juvenile leaves are strong relative to adult leavesand to the leaves of other species. Leaf strength is providedby fibres associated with the midrib. Adult leaves are shorter,broader and less massive than juvenile leaves, and are orientedhorizontally. Transitional leaves are morphologically intermediatebetween juvenile and adult leaves. The anatomy of juvenile,transitional and adult leaves is similar. The heteroblasticseries may be associated with changes in leaf construction costs,light interception and heat dissipation.Copyright 1993, 1999Academic Press Lancewood, Pseudopanax crassifolius (A. Cunn.) C. Koch., leaf heteroblasty, anatomy, morphology, structural adaptations     

Distribution of Lithocysts, Trichomes, Hydathodes and Stomata in Leaves of Pilea cadierei Gagnep. & Guill. (Urticaceae)

In leaves of Pilea cadierei lithocysts, trichomes and hydathodesin the upper epidermis, and lithocysts, trichomes and anomocyticstomata in the lower epidermis are components of a regular distributionpattern in which spacing probably depends upon inhibitory interactionsbetween initial cells during leaf development. Anisocytic stomata,which also occur in the lower epidermis, show a clumped distributionpattern that results from their relatively late formation aftercell division in the epidermis is otherwise complete, and fromthe inhibitory influence of established lithocysts, trichomesand anomocytic stomata. Ordered spacing of anisocytic stomatawithin groups probably results from mutual inhibition betweeninitial cells at the time of inception. Lithocysts associatedwith primary and secondary veins are elongated along the veinaxis. Elsewhere in the leaf lithocyst, orientation is obliquerelative to the midrib and shows a correlation with vein orientationaround the time of lithocysts elongation. Lithocyst, cystolith, trichome, hydathode, stomata, Pilea cadierei     

Effect of Cuticular Abrasion on Thermocouple Psychrometric In Situ Measurement of Leaf Water Potential

Cuticular resistance to water vapour diffusion between the substomatalcavity and the sensing psychrometer junction is a problem uniqueto leaf hygrometry. This resistance is not encountered in soilor solution hygrometry. The cuticular resistance may introduceerror in the measurement of leaf water potential. Using in situleaf hygrometers, we studied the effect of abrading the cuticleof Citrus jambhiri Lushington leaves, to reduce the diffusiveresistance. Field measurements of psychrometer water potentialwere compared with Scholander pressure chamber values for adjacentleaves. Different treatments were compared by sealing pairsof psychrometers on either side of the midrib. The time forwater vapour equilibration between the leaf and the psychrometerchamber was greater than 5 h for no abrasion. For abraded leaves,the true water potential value was obtained within an hour.After equilibration, psychrometer values compared favourablywith pressure chamber values for adjacent leaves (r > 0.97).Measured water potential for unabraded leaves did not correlatewell with corresponding pressure chamber measurements. Scanning electron micrographs indicated that the damage causedby abrading leaves for 60 s using carborundum powder (60 µmdiameter) was surface localized, with numerous scratchings ofthe leaf cuticle. The coarse abrasion treatment (aluminium oxide,75 µm diameter) resulted in fewer but larger cavitiesin the epidermis, which may explain the observed variabilityin the corresponding psychrometric measurements. Key words: Leaf water potential, Cuticular resistance, Leaf abrasion, Thermocouple psychrometer     

The Mechanical Roles of Clasping Leaf Sheaths: Evidence fromArundinaria tecta(Poaceae) Shoots Subjected to Bending and Twisting Forces

Representative shoot segments of the grass speciesArundinariatéctaconsisting of one intact internode and its subtendingnode and clasping leaf sheath were tested to determine the mechanicalinfluence of the leaf sheath on the ability of stems to resistbending and twisting forces. These segments were also used tomeasure shoot morphometry and composite tissue Young's and shearmoduli (EandG,respectively) to simulate the global deformationpatterns attending bending and twisting by means of finite elementanalyses. On average, leaf sheaths contributed 33% of the overallbending stiffness and 43% of the overall torsional stiffnessof stem segments. Comparisons betweenEandGof isolated internodesand leaf sheaths indicated that sheaths were composed of stiffertissues measured either in bending or twisting. Thus, leaf sheathscould act as an external cylindrical brace composed of stiffermaterials than those of the internodes they enveloped. The magnitudesof internodalEandGwere correlated with internodal shape suchthat the ability of internodes to resist twisting relative tothe ability to resist bending forces decreased as internodesbecame more slender or developed thinner walls (both of whichoccur in an acropetal direction from the base to the tip ofshoots). Finite element simulations predicted that, in bending,the leaf sheath laterally braces internodal walls as they tendto ovalize in cross section and push against its inner surfacewhich ovalizes to a lesser extent in the plane normal to thecurvature of shoot flexure. In twisting, the successive ovalizedtransections of internodal walls assumed a helical pattern alongthe length of shoot segments. This helical deformation patternwas attended by an inner lateral contraction of internodal wallsthat was less developed in the leaf sheath that thus provideddecreasing mechanical support to the internode as the lateralcontraction of internodal walls amplified. The twisting of internodesand sheaths was also predicted to concentrate tensile and shearstrains in the nodal diaphragm. Here stress intensities sufficientto produce tissue shear failure were concentrated at two opposingpoints on the surface of the diaphragm. Finite element analysesthus identified a potential weak point in the mechanical constructionof hollow, septate shoots that are, nevertheless, more thanadequately stiff to support their own weight, yet sufficientlyflexible to twist without irreparable damage in normal winds.Copyright1998 Annals of Botany Company Plant stems; nodes; internodes; leaf sheaths; elastic moduli; wind lodging; biomechanics.     

Leaf anatomy of the Pittosporaceae R. Br.

WILKINSON, H. P., 1992. Leaf anatomy of the Pittosporaceae R. Br. An anatomical study of the leaves of 58 species representing all nine genera has been made. The anatomical characters found to be of most use in distinguishing taxa are: in surface view-cuticular architecture as seen with the SEM, stomatal outline, occasionally stomatal density, presence/absence of hairs, hair type in adult leaves; in transverse section-petiole/midrib outline, midrib number of vascular bundles and number of secretory ducts, adaxial epidermis in 1/2 layers, dimensions of adaxial epidermal cells, thickness of outer wall of adaxial epidermis, occasionally chlorenchyma interrupted/not interrupted above the midrib vascular tissue in Pittosporum species; leaf margin.     

Bud formation in leaves,leaf fragments and midrib pieces of Heloniopsis orientalis (Liliaceae)

Summary Isolated leaves, leaf fragments and pieces of the midrib portion devoid of lamina, of Heloniopsis orientalis were grown on an inorganic nutrient medium without organic nutrients and growth regulators in order to investigate their regenerative ability. Bud formation in intact, attached leaves occurs only at the tip, in isolated leaves at the tip and the base, whereas leaf fragments cut transversely at a distance from the tip and isolated midrib pieces form numerous shoot buds in a random distribution. Lamina fragments lacking midrib frequently fail to regenerate even after a long time of culture. It is suggested that endogeneous growth regulators in the leaf, especially the vascular tissues, play an important role in bud initiation. Very young leaves of Heloniopsis are capable forming buds and roots when isolated from the mother plants.     

Xylem cavitation in the leaf of Prunus laurocerasus and its impact on leaf hydraulics

This paper reports how water stress correlates with changes in hydraulic conductivity of stems, leaf midrib, and whole leaves of Prunus laurocerasus. Water stress caused cavitation-induced dysfunction in vessels of P. laurocerasus. Cavitation was detected acoustically by counts of ultrasonic acoustic emissions and by the loss of hydraulic conductivity measured by a vacuum chamber method. Stems and midribs were approximately equally vulnerable to cavitations. Although midribs suffered a 70% loss of hydraulic conductance at leaf water potentials of -1.5 MPa, there was less than a 10% loss of hydraulic conductance in whole leaves. Cutting and sealing the midrib 20 mm from the leaf base caused only a 30% loss of conduction of the whole leaf. A high-pressure flow meter was used to measure conductance of whole leaves and as the leaf was progressively cut back from tip to base. These data were fitted to a model of hydraulic conductance of leaves that explained the above results, i.e. redundancy in hydraulic pathways whereby water can flow around embolized regions in the leaf, makes whole leaves relatively insensitive to significant changes in conductance of the midrib. The onset of cavitation events in P. laurocerasus leaves correlated with the onset of stomatal closure as found recently in studies of other species in our laboratory.     

Effects of Nitrogen on the Development and Growth of the Potato Plant. 1. Leaf Appearance, Expansion Growth, Life Spans of Leaves and Stem Branching

Potatoes (Solanum tuberosum L) were planted in pots in a temperature-controlledglasshouse to collect data on the rate of leaf apearance, leafexpansion, apical lateral branching and active life spans ofleaves The treatments consisted of three rates of nitrogen supply,i e the NI treatment with 2 5 g N per pot and the N2 and N3treatments with 8 and 16 g N per pot, respectively The rate of leaf appearance was 0·53 leaves d^–1(one leaf per 28 °C d) and was negligibly affected by nitrogensupply The rate of leaf expansion was related to leaf numberand nitrogen supply The areas of mature leaves increased withleaf number on the main stem to reach a maximum for leaf numbers12–14, and declined for higher leaf numbers Leaves onapical lateral branches declined in mature area with increasein leaf number The expansion rate of leaves was the dominantfactor that determined the mature leaf area, irrespective ofleaf number and nitrogen treatment The smallest leaves wereobserved at the lowest rate of nitrogen supply Nitrogen promotedapical branching and hence the total number of leaves that appearedon a plant The proportion of total leaf area contributed byleaves on apical branches increased with time and nitrogen supply Active life span, i e the period of time between leaf appearanceand yellowing of the leaf, showed a similar relation to leafnumber as mature leaf area, at least in qualitative terms Leavesof the N3 treatment showed systematically longer life spansthan leaves of the NI and N2 treatment in the order of 3 weeksThe number of main stem leaves was not affected by nitrogensupply Potato, Solanum tuberosum L, leaf development, leaf extension, plant structure, nitrogen nutrition     

The Effect of Temperature on the Growth of Individual Leaves of Vicia faba L. in the Field

The generalized logistic curve was used to describe the growthof individual leaves in crops of Vicia faba L. Durations of.expansionand mean absolute growth rates were derived from these curves.The duration of expansion was inversely related to temperatureaveraged over four days from unfolding. This relationship wasindependent of leaf position except for the lowest leaves. Theduration of expansion of a leaf was related to the rate of productionof new leaves, the number of expanding leaves remaining relativelyconstant. Absolute growth rates varied with leaf position upto leaf 10. At higher leaves, in the absence of water stress,absolute growth rate was a function of temperature and radiation. Vicia faba L., field bean, leaf growth, temperature     

Variation of Bending Capacity Along the Lamina Length of a Grass, Imperata cylindrica var. major (Gramineae)

The aim of this paper is to investigate variation of the bendingmoment capacity along the lamina of a grass (Imperata cylindrica(L.) Raeusch. var. major (Nees) C. E.Hubb. ex C. E.Hubb. &Vaughan). It is postulated that the bending strength of thegrass section is obtained predominantly from the midrib andthat the grass lamina contributes little to the overall bendingbehaviour. By observing transverse sections of the midrib, itcan be estimated that the thickness of the outer sclerenchymazone is approx. 10% of the midrib diameter and that the pithparenchyma makes up most of the rest of the midrib. This observationis taken to be valid for the entire length of the lamina. Asimple structural model of an elliptical tube is proposed forthe midrib. Bending tests on the grass lamina have shown thatthe bending moment capacity is very low in the distal third,but increases linearly in the middle third and is generallyconstant in the proximal third. Using the structural model forthe midrib, the results indicate that the maximum bending stressof the midrib remains fairly constant over the length of thegrass. Copyright 1999 Annals of Botany Company Bending strength, maximum moment capacity, grass, Imperata cylindrica.     

Leaf Ecology of Pre-reproductive Ontogenetic Stages of the Palm TreeEuterpe edulisMart. (Arecaceae)

We studied leaves of 208 seedlings (S), infants (I) and juveniles(J) ofEuterpe edulis,randomly selected from plants in 1 ha ofswampy forest in SE Brazil. Each new leaf began extending afterthe complete development of the preceding leaf. The sequencesteps of leaf growth were emergence, linear growth while closed,opening of segments and logarithmic growth of the petiole. Averageleaf production rate (2.21 leaves per plant year^-1) did notvary among ontogenetic stages, conforming to a plastochronicrhythm. Average linear growth rate of the closed leaf was directlyrelated to average area of the open lamina. Herbivores attacked33.0% of all leaves giving rise to up to 10% loss of area andincluded damping-off-inducing suckers (7.1%), ordinary suckers(9.8%), chewers (5.4%), miners (2.7%) and multiple attacks (8.0%).These attacks varied among stages (S=22.4%, I=38.1%, J=33.3%),as did leaf mortality rates (S=26.5%, I=14.3% and J=0.0%). Unknownfactors were the main cause of leaf mortality (S=14.3%, I=7.1%).Not losing a leaf while a seedling and attaining a minimum leafarea in the infant stage were critical events for survival.Plants gained leaf area by not losing leaves while a seedling,by producing larger leaves with greater growth rates, and byaccumulating leaves with longer lifespans. The petiole can simulatean energetically cheaper branch, delimit a vital space aroundthe stem, favourably position the leaf lamina and substituteprovisionally for stem growth in height.Copyright 1999 Annalsof Botany Company Arecaceae,Euterpe edulis, herbivory, leaf ecology, leaf growth, leaf production, semideciduous forest, southeastern Brazil.     

Branching Principles Governing the Architecture of Cornus kousa (Cornaceae)

The complex structure of the crown of Cornus kousa, generallyfive-forked in vegetative branching and two-forked in reproductivebranching, is analysed quantitatively and described by two basicbranching principles: decussate phyllotaxy and the resettingrule for planes of branching. Most Cornus species have opposite,decussate phyllotaxis. The leaf pair (with axillary buds) definesthe branching plane of a node. Because of regular phyllotaxis,the fundamental branching pattern is that every branching planealong an axis is perpendicular to the preceding one. However,the first node of a lateral horizontal shoot always has a horizontalbranching plane; we term this the resetting rule. We observedthat resetting occurs when the first nodes initiated in thevertical plane are repositioned by a twisting of their firstinternodes. All later nodes alternate directions, i.e. showusual decussate alternation. Foliage leaf nodes usually producethree-forked branchings. When vegetative winter buds are formed,a foliar node and adjacent scale leaf node produce a five-forkedbranching. When reproductive winter buds with a terminal inflorescenceare formed, the last foliar node and two adjacent scale leafnodes can produce a variety of branchings but usually producean equal two-forked branching. To understand better the architecturein C. kousa, we contrast it with C. capitata which does notproduce buds with scale leaves and whose vegetative nodes areclearly separated. Copyright 1999 Annals of Botany Company Branching pattern, Cornaceae, Cornus kousa, decussate branching, dogwood, Japanese strawberry tree, tree architecture, tree geometry.     

Senescence in Leaves of Acer pseudoplatanus L. and Parthenocissus tricuspidata Planch: 1. Changes in Some Leaf Constituents during Maturity and Senescence

Data are recorded of the changes in chlorophylls, carotene,sugar, shikimic acid, and anthocyanin in leaves of sycamore(Acer pseudoplatanus L.) and Virginia creeper (Parthenocissustricuspidata Planch.) during leaf maturity and senescence. InParthenocissus the losses of chlorophylls, carotene, sugar,and shikimic acid during senescence were closely correlated,and were inversely related to the accumulation of anthocyanin.The losses of chlorophylls, carotene, and sugar by Acer leaveswere also closely correlated. No evidence was found to supportthe suggestion that anthocyanin formation was caused by accumulationof sugar during senescence, but marked differences in shikimicacid content were found between leaves of Acer, which did notform anthocyanin, and Parthenocissus. which did. It is suggestedthat autumn senescence of these leaves involves the rapid senescenceof an increasing proportion of the leaf tissue during a periodof 80 days, and that measurements of the content of constituentsgive an estimate of the proportion of the leaf tissue whichhas senesced.     

Seasonal Dimorphism in the Mediterranean Cistus incanus L. subsp. incanus

Mediterranean perennial species are described as being sclerophyllous,or summer deciduous, or seasonally dimorphic. Field observationin the coastal maquis of Castelvolturno Nature Reserve, southernItaly, showed thatCistus incanus L. subsp. incanus is a seasonallydimorphic species as it develops brachyblasts with small leavesin summer, and dolichoblasts with large leaves in winter. Fieldbiometric data confirmed that winter shoots were 14-times longerthan those developed in summer and had many more leaves. Thearea of single winter leaves was five-times that of summer leaves.Anatomical leaf structure also changed with the season: winterleaves were flat while summer leaves had a crimped lamina whichwas partially rolled to form crypts in the lower surface. Leaveswere covered by considerably more trichomes in summer than inwinter. Stomata were uniformly distributed along the lower epidermisof winter leaves but were only present in the crypts of summerleaves. In summer leaves, a palisade layer was often found onboth sides of the lamina, the mesophyll cells were generallysmaller and the intercellular spaces were reduced. Winter leaveshad a dorsiventral structure and larger intercellular spaces.Seasonal dimorphism is generally reported to be an adaptationto summer drought. However, the morphology and anatomy of C.incanus L. subsp.incanus showed that the subspecies has notonly developed a strategy to survive summer drought, but hasevolved two different habits, one more xerophytic than the other,to optimize adaptation to the seasonal climatic changes occurringin Mediterranean environments. Copyright 2001 Annals of BotanyCompany Cistus, Cistus incanus L. subsp. incanus, climatic changes, leaf anatomy, leaf dimorphism, Mediterranean shrubs, phenology, seasonal dimorphism     

Abscisic Acid and the Regulation of Water Loss in Plantlets of Brassica oleracea L. var. botrytisRegenerated Through Apical Meristem Culture

Water loss was studied in regenerated plantlets of Brassicaoleracea var. botrytis cv. Currawong derived through apicalmeristem culture. Hardening of plantlets was eliminated by asingle application of a polyvinyl resin (S600) sprayed immediatelyafter transplanting. Plantlets sprayed with S600 had highercuticular resistances than unsprayed plantlets; this treatmenthad no effect on stomatal resistance. Leaves formed during theculture period showed very little wax formation and using markedleaves it was found that only reduced levels of wax formed onthese leaves even after transplanting. New leaves formed aftertransplanting, showed typical wax formation compared to seedgrown plants. Abscisic acid (ABA) at 10^–4 M applied as a leaf sprayto transplants did not cause a substantial increase in stomatalresistance in leaves which had been initiated during the cultureperiod. Leaves of seed-grown plants as well as leaves of plantletsformed after transplanting did respond to a leaf spray of ABAat 10^–4 M by a large increase in stomatal resistance. Relative concentrations of K, Na, Ca, P, S and Mg in guard cellswere calculated for each leaf type by X-ray micro-probe analysis.K/Na values decreased in the order: seedling > leaves formedafter transplanting > leaves intiated during culture. A highpositive correlation was also found between K/Na and K/P forthe three leaf types. K:Mg and K:Ca ratios for leaves formedduring culture were low in comparison to the values obtainedfor leaves formed after transplanting and seedlings for whichthe values were similar. Brassica oleracea var. botrytis, cauliflower, regenerated plantlets, meristem culture, stomatal resistance, water loss, abscisic acid, X-ray micro-probe analysis     

Spatial distribution characteristics of stomata at the areole level in Michelia cavaleriei var. platypetala (Magnoliaceae)

Background and AimsIn hierarchically reticulate venation patterns, smaller orders of veins form areoles in which stomata are located. This study aimed to quantify the spatial relationship among stomata at the areole level.MethodsFor each of 12 leaves of M. cavaleriei var. platypetala, we assumed that stomatal characteristics were symmetrical on either side of the midrib, and divided the leaf surface on one side of the midrib into six layers equidistantly spaced along the apical–basal axis. We then further divided each layer into three positions equidistantly spaced from midrib to leaf margin, resulting in a total of 18 sampling locations. In addition, for 60 leaves, we sampled three positions from midrib to margin within only the widest layer of the leaf. Stomatal density and mean nearest neighbour distance (MNND) were calculated for each section. A replicated spatial point pattern approach quantified stomatal spatial relationships at different distances (0–300 μm).Key ResultsA tendency towards regular arrangement (inhibition as opposed to attraction or clustering) was observed between stomatal centres at distances <100 μm. Leaf layer (leaf length dimension) had no significant effect on local stomatal density, MNND or the spatial distribution characteristics of stomatal centres. In addition, we did not find greater inhibition at the centre of areoles, and in positions farther from the midrib.ConclusionsSpatial inhibition might be caused by the one-cell-spacing rule, resulting in more regular arrangement of stomata, and it was found to exist at distances up to ~100 μm. This work implies that leaf hydraulic architecture, consisting of both vascular and mesophyll properties, is sufficient to prevent important spatial variability in water supply at the areole level.