Stomatal Features in the Leaf of Aganosma dichotoma (Roth) K. Schum

Paracytic and anisocytic types of mature stomata are found inthe leaf of Aganosma dichotoma. Stomata with one guard cell,stomata with degenerated guard cells, and contiguous stomataare common. Stomata with arrested pore development are alsofound in certain cases. A single guard cell without any porehas not been designated as a stoma with one guard cell in thepresent investigation. Ontogeny of contiguous stomata have beentraced. Subsidiary cells are, morphologically, just like theircontiguous guard cells. Subsidiary cells may retain their shapeand contents even when their contiguous stoma becomes mature,or may change their shape and lose their contents. They mayor may not divide. Subsidiary cells form a whorl of more thantwo subsidiary cells around a stoma by their divisions. Degenerationof guard cell(s)— their contents and nuclei—havebeen traced. In certain cases guard cells divide forming morethan two guard cells associated to a single pore. Cytoplasmicconnections are found between two guard cells of nearby stomata,and between a guard cell and an epidermal cell. Near the wound,the epidermal cells over the veins become meristermatic givingrise to new epidermal cells but no meristemoid.     

Variable Cell Lineages form the Functional Pea Epidermis

Evidence was sought for cellular programs and cellular interactionsacting during the formation of stomatal spacing patterns. Dailyreplicas of the surfaces of Pisum sativum leaves were used toreconstruct the cellular development of specific regions ofthe epidermis. During the period studied small primordia becamemature leaves; this involved a 250-fold increase in area anda 20-fold increase in cell number. The earliest event correlatedwith the development of a stoma was an unequal division, andsuch divisions were common in neighbouring and even within thesame cells. A distinct cell lineage started with these unequaldivisions, forming both a stoma and most of the cells that separatedit from its neighbours. Both products of an unequal divisionbecame regular epidermal cells only where such development preventedthe formation of two stomata that would have been in directcontact with one another. Neighbouring stomata often developedand matured together, indicating that there was no mutual inhibitionbetween developing stomata that were more than one cell apart.It is concluded that stomata are products of an intracellularprogram which generates stomatal patterns during rather thanpreceding development. This program can be modified and evenstopped during its entire course, allowing for the correctionof local ‘mistakes’ of stomatal patterning. Cell lineages, cell determination, cellular interactions, epidermal development, garden peas, immature stomata, pattern formation, Pisum sativum, spacing patterns, stomata, unequal divisions     

Metabolic Inhibitors Block ABA-Induced Stomatal Closure

Closure of stomata of Commelina communis L. leaf epidermis causedby abscisic acid (ABA) was inhibited by sodium azide, potassiumcyanide and hypoxic conditions. Azide was more effective thancyanide at low concentrations, but the cyanide effect couldbe enhanced by addition of salicylhydroxamic acid, providingindirect evidence for cyanide-resistant respiration in epidermaltissue. Azide also inhibited ABA-induced closure of ‘isolated’ stomata and shrinkage of guard cell protoplasts.The results indicate that metabolic energy is required for ABAaction involving solute loss from the guard cells. Possiblemechanisms of action are discussed.     

The development of stomata and other epidermal cells on the rice leaves

In the leaves of rice (Oryza sativa), stomatal initials arose from two asymmetric cell divisions and a symmetric division. Guard mother cells (GMCs) and long cells in stomatal files (LCSs) were formed through the first asymmetric division of the precursor cell of GMCs. Subsidiary cells (SCs) were produced by the second asymmetric division of subsidiary mother cells or LCSs. Following SC formation, GMCs divided once symmetrically to generate guard cells and then differentiated terminally to form mature stomata. The developmental patterns of long cells, prickle hairs and short cells (phellem cells and silica cells) were also examined. Interestingly, we found that the different developmental stages of stomata and epidermal cells occurred in the similar location of immature leaves of the same phyllotaxis. In addition, two spacing patterns (“one stoma, one long cell” and “one short cell row”) probably exist in rice leaves.     

Developmental mechanism and distribution pattern of stomatal clusters in Cinnamomum camphora

We report the stomatal cluster development mechanism and distribution pattern in Cinnamomum camphora. The results indicated that the clustered arrangement of meristemoids at the juvenile stage of the leaf development contributed greatly to the pattern of stomatal clusters. Additionally, division of an epidermal cell (EC), which is between small stomata, and growth of small stomata to push the ECs aside to become directly contacting had an impact on the development as well as the pattern of stomatal clusters. The latter way may play a more important role in stomata clustering of the wide-type C. camphora. There are no significant difference in the stomatal index (the number of stomata per the sum of the number of epidermal cells and the number of stomata) among different part of leaves, while the stomatal cluster index (the number of stomata in stomatal clusters per the total number of stomata) was found to increase gradually from the apex to the base and from the middle part to the marginal part of the leaf. The possible reason of this pattern was discussed. Published in Russian in Fiziologiya Rastenii, 2006, Vol. 53, No. 3, pp. 348–353. The text was submitted by the authors in English.     

Responses of Commelina communis Stomata In Vitro

Weyers, J. D. B. and Paterson, N. W. 1987. Responses of Commelinacommunis stomata in vitro.— J. exp. Bot. 38: 631–641. Analysis of the kinetics of movements of Commelina communisL. stomata in vitro revealed a sequence of opening and closingphases dependent on the incubation medium used and the physiologicalstate of the plant material. In buffer containing 50 mol m ^–3KC1 the sequence of aperture changes appeared to be influencedby equilibration of cell water potentials with that of the mediumand by solute fluxes (dependent and independent on metabolicactivity). The results indicate that the stomatal aperture afterseveral hours of incubation may not always provide a reliablequantitative estimate of the ability of the stomata to operate.As a consequence, modifications are suggested to the ways inwhich experiments using epidermal strips are carried out andreported. Key words: Epidermal strips, guard cells, hydroactive, hydropassive, kinetics, potassium chloride, mannitol, osmotic effects, solute fluxes.     

The Structure and Orientation of Guard Cells in Plants Showing Stomatal Responses to Changing Vapour Pressure Difference

Transmission electron micrographs revealed that a substantialpart of the guard cell wall of both Quercus robur L. and Populusnigra ‘italica’ L. was either free of cuticle orcovered with a greatly reduced cuticular layer. In Quercus thestructure of the guard cell was such that the area of limitedcuticular development would be exposed to the evaporating powerof the atmosphere even when the stomata were closed. Lanthanumstaining confirmed that this area might be an important siteof evaporation. A similar evaporation site was identified inthe guard cell wall of Pinus sylvestris L. Light micrographsrevealed that this area could also be exposed on the outsideof the leaf when the stomata were closed. It appears that guardcell orientation with respect to the epidermal plane dependsupon epidermal turgor. Changes in orientation of the guard cellcoupled with the exact location of the cuticle-free area inthe guard cell wall may explain the nature of the stomatal responseof individual species to changing VPD and the effect of othervariables, e.g. water deficit, on this response. Quercus robur L, oak, Populus nigra L, poplar, stomata, guard cells, cuticle, evaporation, vapour pressure difference     

Stomata and Structure of Tetraploid Apple Leaves cultured in Vitro

Leaves of anther-derived tetraploid apple (Malus pumila Mill.)shoots were examined by low-temperature scanning electron microscopy(LT-SEM). Leaves were serrate and wide with an undulating adaxialsurface due to convex epidermal cells, apparently without crystallineepicuticular wax. Stomata were absent from the adaxial surface,except for the marginal teeth which exhibited 40-60 stomataper leaf; they probably originated from residual mitotic activity.One third of abaxial stomata was occluded by the residual cuticleof the mother guard cell across the stomatal pore which rupturedwhen the stomata became functional. The stomatal index was 7·2(± 1·6) with 60-75 stomata mm^-2, i.e. abaxialstomata of tetraploid leaves expanded in vitro were less frequentthan those in triploid leaves either cultured in vitro (475-575stomata mm^-2) or grown on the tree (320-390 stomata mm^-2) wherethe stomatal index was 21 (± 4). Freeze-fracture transsectionsshowed that the tetraploid in vitro leaves were composed ofa layer of adaxial epidermal cells, followed by a single layerof palisade cells and four to five layers of spongy mesophyllcells and the abaxial layer of epidermal cells, in contrastto juvenile seedling-grown apple leaves in which the two layersof palisade cells comprised the majority (52-60%) of the leafvolume. The same morphological features, such as wide and lesspointed leaves, reduced stomatal density and stomatal index,and increased stomatal size that were previously reported fortree-grown tetraploid leaves were also expressed in vitro. Thus,causes of the stomatal deformation in tissue-cultured Rosaceaeare interpreted to be in part genetic and not purely environmental.Copyright1994, 1999 Academic Press Malus pumila Mill., apple, biotechnology, breeding, cryo-preservation, CO₂, juvenile, low temperature-scanning electron microscopy (LT-SEM), micropropagation, ploidy, stomata, tissue-culture, transpiration     

Epidermal Structures and Stomatal Ontogeny in Some Mimosaceae

The structure of trichomes and stomata on leaflets of 21 speciesof the Mimosaceae are described. Non-glandular trichomes inMimosa pudica are of three types: unicellular, with a roundedthick-walled base and a terminal unicellular body, and multicellular.Capitate, clavate, or cylindric, 3–6-celled glandularhairs are observed on leaflets of Mimosa pudica only. Leafletsare amphistomatic in all species except Adenanthera pavonina,Calliandra sp., Parkia biglandulosa, Pithecellobium dulce, andSamanea saman in which they are hypostomatic. Only paracyticstomata are found in Leucaena leucocephala and Mimosa pudica.In the rest stomata are of more than one type. In spite of thediversity, the most frequent type in these species is paracytic.Anisocytic stomata, in all cases, are secondarily derived fromparacytic ones by transverse or oblique wall formation in asubsidiary cell. Similarly some stomata with one subsidiarycell are also secondary derivatives of the paracytic ones becauseof one of the subsidiary cells assuming the form of an epidermalcell. The development has been traced in 14 species and thatof paracytic stomata may be mesogenous or mesoperigenous, thatof stomata with one subsidiary cell mesogenous but anomocyticstomata are ontogenetically perigenous. Occasionally a meristernoidis cut off from one of the subsidiary cells of a paracytic stoma.The organization of a stoma from such a meristemoid has beentraced.     

Stomatal patterning in Tradescantia: an evaluation of the cell lineage theory.

The cell lineage theory, which explains stomatal patterning in monocot leaves as a consequence of orderly divisions, was studied in Tradescantia. Data were collected to test the theory at three levels of organization: the individual stoma; stomata distributed in one dimension, in linear fashion along cell files; and stomata apportioned in two dimensions, across the length and breadth of the leaf. In an attempt to watch the patterning process through regeneration, stomata in all visible stages of development were laser ablated. The results showed that the formation of stomatal initials was highly regular, and measurements of stomatal frequency and spacing showed that pattern was determined near the basal meristem when the stomatal initials arose. Following the origin of initials, the pattern was not readjusted by division of epidermal cells. Stomatal initials were not committed when first present and a small percentage of them arrested. The arrested cells, unlike stomata, were consistently positioned in cell files midway between a developed pair of stomata. At the one-dimensional level of pattern, stomata in longitudinal files were separated by a variable number of epidermal cells and the frequency of these separations was not random. The sequential spacing of stomata also was not random, and stomata separated by single epidermal cells were grouped into more short and long series than expected by chance. The stomatal pattern across the width of the leaf resulted from cell files free of stomata which alternated with cell files containing stomata, but not with a recurring periodicity. Files lacking stomata were found only over longitudinal vascular bundles. Laser ablations of developing stomata did not disrupt the pattern in nearby cells or result in stomatal regeneration. We conclude that the cell lineage theory explains pattern as an individual stomatal initial arises from its immediate precursor and satisfactorily accounts for the minimum spacing of stomata in a cell file, i.e., stoma-epidermal cell-stoma. However, the theory does not explain the collective stomatal pattern along the cell files, at the one-dimensional level of patterning. Nor does the theory account for the for the two-dimensional distribution of stomata in which regions devoid of stomata alternate with regions enriched with stomata, but not in a highly regular nor haphazard manner. We suggest that the grouping of epidermal cells and stomata separated by single epidermal cells in cell files may result from cell lineages at a specific position in the cell cycle as they traverse the zone where stomatal initials form.(ABSTRACT TRUNCATED AT 400 WORDS)     

Anatomy and Transpiration of the Avocado Inflorescence

Structure and function of the inflorescence of cv. 'Hass' and'Fuerte' avocado (Persea americana Mill.) were examined by scanningelectron microcopy (SEM) and by porometry. Sepals and petalscould not be distinguished by their position in the flower,by visual gross morphology or by microscopic surface structureand were hence designated as tepals. These tepals were arrangedin two whorls of three, followed by two whorls of three outerand three inner stamens, each opposite a tepal. The most conspicuousfeature of tepals, developing leaves and peduncles was the densecover of hair which were most frequent on the adaxial tepalsurface (925-1200 trichomes mm^-2), followed by their abaxialsurface (625-1000 mm^-2) and peduncles (375-655 mm^-2). Stomatawere absent from the adaxial surfaces of both tepal and leaves,as well as peduncles. On the tepals, abaxial stomata appearedfunctional, small (8-9 x 11-13 µm) and scarce with 2·8-3·4stomata mm^-2, i.e. very low relative to avocado leaves (350-510stomata mm^-2) or young fruit (50-75 stomata mm^-2. However, flowersincluding tepals transpired 1·2-1·3 mmol underfield conditions in Southern California (1·6-2 kPa),i.e. in excess of leaves (0·7-1·1 mmol) and peduncles(0·6-0·8 mmolH₂O m^-2 s^-1). This situation wasattributed to the few small but functional abaxial stomata onthe tepal, in contrast to 80% closed stomata and dense epicuticularwax cover in form of rodlets on young and dendritic crystalson old leaves including the guard cells, and absence of stomatafrom the peduncle.Copyright 1993, 1999 Academic Press Persea americana Mill., avocado, bioenergetics, flower, fruit, leaf, peduncle, scanning electron microscopy, stomata, transpiration, petals, sepals, tepals     

Epidermal Structure and Development of Stomata in some Annonaceae

Epidermal structure and development of stomata are studied in20 species of the Annonaceae. Epidermal cells are polygonalor irregular in outline, isodiametnc or elongated with straight,arched, or sinuous anticlinal walls. Cuticular striations radiatingfrom guard cells or hair bases are noticed. Four types of trichomesand secretory cells are seen. The mature stomata are paracyticwith 2–6 subsidiary cells, non-contiguous at both poles,or contiguous at one or both the poles. The increase in numberof subsidiary cells is due to their divisions. Contiguous stomata,arrested development, single guard cell, and degeneration ofguard cells are observed. The development is either syndetocheilicor mesogenous.     

Electrical Currents at the Leaf Surface of Commelina communis and their Relationship to Stomatal Activity

A vibrating probe was used to detect and measure electricalcurrents at the surface of excised leaves and isolated leafepidermis from Commelina commnunis. Currents of up to 4.0 µAcm^–2 moving out from the leaf surface were observed whenthe stomata were open. When the stomata were almost closed nocurrent was detected and when they were fully closed a smallinwardly directed current was observed. There appeared to bea linear relationship between current and stomatal aperture.The current was stimulated by fusicoccin and eliminated by increasingthe external pH suggesting that it was brought about by a flowof H⁺ from the leaf surface. Key words: Electrical currents, leaves, stomata, vibrating probe     

Role of topography sensing for infection-structure differentiation in cereal rust fungi

Over 90% of the germ tubes of Puccinia graminis tritici (wheat stem rust) and Puccinia hordei (barley brown rust) differentiate appressoria on encountering stomata.There has been controversy as to the role of host topographical signals in the highly precise and efficient induction of these infection structures over stomata by cereal rusts. In the present study, polystyrene replicas of microfabricated silicon wafers, bearing precise microtopographies of defined dimensions, were used to investigate the influence of ridge spacing and height on infection-structure induction by P. graminis tritici and P. hordei. It was found that artificial topographical signals alone can induce a reproducibly high percentage (83–86%) of germ tubes to differentiate infection structures. Multiple, closely spaced (1.5 μm) ridges which were 2.0 μm high provided the most inductive topography. Differentiation on flat surfaces and over single ridges was < 4%. Appressorium induction commonly initiated a cascade of differentiation events involving the formation of infection pegs, vesicles, infection hyphae, and occasionally haustorial mother cells. It is suggested that the close spacing of cell junctions associated with the dumbbell-shaped guard cells of cereal stomatal complexes provide inductive signals for infection-structure formation by cereal rusts in vivo. Received: 17 October 1996 / Accepted: 5 December 1996     

Variation in the structure and development of foliar stomata in the Euphorbiaceae

The structure and ontogeny of foliar stomata were studied in 50 species of 28 genera belonging to 17 tribes of the family Euphorbiaceae. The epidermal cells are either polygonal, trapezoidal, or variously elongated in different directions and diffusely arranged. The epidermal anticlinal walls are either straight, arched or sinuous. The architecture of cuticular striations varies with species. The mature stomata are paracytic (most common), anisocytic, anomocytic and diacytic. Occasionally a stoma may be tetracytic, cyclocytic or with a single subsidiary cell. The ontogeny of paracytic stomata is mesogenous dolabrate or trilabrate, mesoperigenous dolabrate; that of diacytic stomata is mesogenous dolabrate, whereas that of anisocytic stomata is mesogenous trilabrate; rarely an anisocytic stoma may be mesoperigenous. Hemiparacytic stomata are mesoperigenous unilabrate; tetracytic stomata are mesoperigenous dolabrate and anomocytic stomata perigenous. Abnormalities encountered include four types of contiguous stomata, stomata with a single or both guard cells aborted and persistent stomatal initials. Cytoplasmic connections between the guard cells of two adjacent stomata or the guard cell of a stoma and an adjacent epidermal/subsidiary cell, or both types occurring in a species, were noticed. The stomatal development, distribution, diversity and basic stomatal type with reference to systematics are discussed.     

Divergent regulation of stomatal initiation and patterning in organ and suborgan regions of the Arabidopsis mutants too many mouths and four lips

Stomata are consistently patterned so that they are not in contact. This patterning is violated in the too many mouths (tmm) and four lips (flp) mutations of Arabidopsis thaliana (L.) Heynh. which have stomatal clusters in the first-formed leaves. To clarify the function of both genes in stomatal initiation and patterning, the phenotypes of many different organs were quantified. The flp mutation affects dorsiventral and cylindrical organs differentially with respect to the frequency of clustering. The tmm mutation has a more complex region-specific phenotype in that some regions lack stomata entirely, other regions have excess stomata, and the flower stalk exhibits an apex-to-base gradient from excess to no stomata. This suggests that TMM represents an unusual type of gene regulating plant cell development in that it can either influence stomatal initiation in a positive or negative fashion depending on region. Since the frequencies of initiation and clustering can be uncoupled in tmm, these two functions are under separate region-specific control. Analysis of double mutants shows that tmm and flp in some cases show region-specific interactions in both cluster formation and initiation, and that there may be subpopulations of stomata under different genetic control. Received: 10 November 1997 / Accepted: 29 December 1997     

Leaf anatomy of highbush blueberry grownin vitro and during acclimatization toex vitro conditions

Leaves of micropropagated highbush blueberry (Vaccinium corymbosum) cv. ‘Bluetta’ have been observed during the acclimatization phase. In vitro-developed leaf cells were circular and small, the spongy parenchyma was discontinuous and disorganized and formed by 1–2 layers of cells with large intercellular spaces and the palisade to spongy mesophyll thickness ratio was 1:1.5. After rooting ex vitro, the first leaves formed under natural conditions showed substantial changes in the anatomical characteristics. After 6 months, the plants produced leaves similar to those in field-grown plants. The palisade cells were rectangular, the spongy parenchyma was formed by 3–4 layers of cells and the intercellulars were around the stomata. Leaves from field-grown plants lost 24 % of water during 150 min after excision while leaves from in vitro shoots lost about 50 % of water in the same time. Leaves from in vitro shoots showed a higher number of smaller stomata (361 per mm²), with the guard cells forming a circular ring; the stomata frequency in field-grown leaves was 241 per mm² and the guard-cells were elliptical.     

Stomata in some species of genus <Emphasis Type="Italic">Arum</Emphasis> from the Eastern Slavonia and Baranya region

Types and the number of stomata in the following Arum species: Arum italicum Mill., Arum maculatum var. maculatum L. and Arum maculatum var. immaculatum L., Arum alpinum var. pannonicum Terpo. and Arum alpinum var. intermedium Schur. in three different locations in Zablaće, Normanci and Bilje were investigated. The most prevalent stomata type at both upper and lower epidermis for each Arum species was mostly stomata type paracytic, followed by hexacytic, tetracytic or brachyparacytic as far as locality is concerned. Helicocytic type was more prevalent in Arum alpinum var. pannonicum Terpo. and Arum alpinum var. intermedium Schur. at Bilje. Other stomata types were very rare. A striking regularity in the occurrence of stomata types was not found within a single species. Some stomata types, however, were found either at adaxial or abaxial epidermis or were not present at all. The number of stomata per square mm varied from 25 to 651. A statistically significant difference in the number of stomata per square mm at upper and lower epidermis among Arum species was determined in locations Zablaće and Normanci, whereas no statistically significant differences were found in location Bilje.     

The Effects of Temperature and ABA on Stomata of Zea mays L.

Epidermal fragments were removed from maize leaves by tearingparallel to the veins. These were incubated at a number of differenttemperatures in several concentrations of ABA. The sensitivityof stomata to temperature was dependent upon the technique usedto incubate epidermis. Generally, the widest apertures wererecorded at around 25°C. In all experiments, stomata incubatedat low (10°C) temperatures on 5.6 x 10^–4 M ABA showedwider apertures than those incubated on distilled water. ThisABA-stimulated stomatal opening was accompanied by an increasein the intensity of potassium staining in the guard cells. At25 °C, epidermis incubated on several concentrations ofABA showed some stomatal closure, decreased potassium stainingin the guard cells and increased potassium staining in the subsidiarycells.     

Effect of Elevated CO2 on Stomatal Size and Distribution in Perennial Ryegrass

Plants of ryegrass (Lolium perenne L. cv. Melle) were grownfrom the early seedling stage in growth cabinets at a day/nighttemperature of 20/15 °C, with a 12-h photoperiod, and aCO₂ concentration of either 340 or 680 ± 15 µl1^–1 CO₂. Young, fully-expanded, acclimated leaves fromprimary branches were sampled for length of stomata, and ofepidermal cells between stomata, numbers of stomata and epidermalcells per unit length of stomatal row, numbers of stomatal rowsacross the leaf and numbers of stomatal rows between adjacentvein ridges. Elevated CO₂ had no significant effect on any ofthe measured parameters. Elevated CO₂, Lolium perenne, ryegrass, stomatal distribution, stomatal size