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.     

中国桑寄生科植物叶表皮微形态

通过扫描电镜对中国桑寄生科桑寄生亚科8属18种和槲寄生亚科1属2种植物成熟叶的上、下叶表皮内表面和下表皮外表面进行了研究。内面观发现桑寄生科植物叶上、下表皮形状为多边形,垂周壁式样平直或稍弓形,常具有角质增厚,平周壁常覆盖厚角质或颗粒状、丝状角质增厚;气孔存在于上下表皮,通常下表皮较多,气孔的形状,特别是保卫细胞的形态在亚科间、属间或种间都具有一定的差异,气孔器类型为平列型或单圈型。下表皮表面观察了的角质膜和蜡质纹饰、气孔的形状,外部气孔缘及外部气孔缘内缘的特征。这些特征在亚科或属级水平上较为稳定,有的也表现出种间差异,有一定的分类价值。从气孔形态和外部气孔周围角质膜来看,两亚科显示出明显的不同:桑寄生亚科上、下表皮均具有内部气孔缘,而槲寄生亚科没有此结构;桑寄生亚科外部气孔周围角质膜增厚成环状,其上具增厚的条纹,而槲寄生亚科外部气孔周围角质膜增厚成脊状,不具条纹。这些特征支持槲寄生亚科作为独立1个科来处理。     

Cuticle micromorphology of leaves of Pinus (Pinaceae) from Mexico and Central America

Cuticle micromorphology of 34 taxa of Pinus from Mexico and Central America was studied with scanning electron microscopy, and leaf morphology was described. In total, 29 characters, 22 from the inner cuticular surfaces and seven from the outer, were described in detail. These characters have value either for testing infragenerie classifications or for identifying individual taxa. Characters relating to the periclinal wall texture of the epidermal cells, the shape and degree of development of the anticlinal walls of the epidermal cells, the basal and apical shapes of anticlinal epidermal cell walls, the continuity of the epidermal cells, the size ratio of the polar to lateral subsidiary cells, the grooves on subsidiary cells, the cuticular flanges between guard and subsidiary cells, the groove near the bristles and the elevation of the Florin ring ridge and striations on the Florin ring are particularly useful for infrageneric classification. The agreement between these characters and infrageneric classifications is discussed. Characters relating to the end wall shapes of the epidermal cells, the relative length of epidermal cells, the shape of the stomatal apparatus, the texture of guard and lateral subsidiary cell surfaces, the polar extensions, the number of subsidiary cells and epidermal cell layers between stomatal rows, the integrity of stomatal rows, cell numbers between stomata in a row, cuticular flanges between guard cells, bristle flanges and surface textures, epicuticular waxes, striations on Florin rings and stomatal shapes, contain some important information for identifying Mexican pines. The distribution of the states of each character is compared with that of the Asian pines. Cuticular characters are used to help determine the affinities of taxonomically difficult taxa.     

Morphological characters of leaf epidermis in schisandraceae and their systematic significance

The leaf epidermis of 23 species belonging to 2 genera within Schisandraceae was investigated using light and scanning electron microscopy. Many characters of the leaf epidermis in Schisandraceae, such as shape of epidermal cells, type of stomata, and cuticular ornamentation, are usually constant within species and thus helpful for elucidating the relationship between and within genera. Leaf epidermal cells are usually irregular or polygonal in shape. The patterns of anticlinal walls are straight, sinuolate, sinuous or sinuate. The stomatal apparatus belong to paracytic or laterocytic type and the latter is subdivided into various subtypes based on the number and arrangement of subsidiary cells. Under scanning electron microscopy observation, the cuticular membrane is often striated, sometimes squamulate or granular; the inner margin of the outer stomatal rim is nearly smooth or denticulate. Evidences from shape of epidermal cells, patterns of cuticular intrusions between the ends of each guard cell of a pair and distribution of stomatal apparatuses support the viewpoint thatKadsura is more primitive thanSchisandra. Study on leaf epidermis also shows thatKadsura interior deserves the rank of a distinct species and the treatment of the evergreen groups, includingS. propinqua andS. plena, as distinct from the deciduous species of the genus is quite natural.     

叶子花表皮特征的研究及意义

对叶子花（Bougainvillea spectabilis）正常叶和变态叶上气孔密度、气孔指数和保卫细胞大小进行了研究。结果表明：正常叶上表皮的表皮细胞为多边形,垂周壁平直;下表皮的表皮细胞为不规则型,垂周壁浅波状;气孔类型为不规则型。变态叶上表皮没有发现气孔,变态叶下表皮的表皮细胞垂周壁则由浅波形逐渐变为深波形,气孔类型为不规则型和轮列型。随着变态叶的发育,变态叶下表皮的气孔密度降低,气孔指数升高;变态叶保卫细胞的长增大,宽减小。变态叶的平均气孔密度和平均气孔指数明显低于正常叶。正常叶和变态叶的保卫细胞均呈肾形。     

The pattern of cuticular blue-fluorescing phenolics deposition during the development of Prunus persica leaves

Although stomatal ontogeny is closely related to the development and maturation of the epidermal tissue, stomatal patterns in relation to cuticle construction and cuticular material deposition during leaf development have not received adequate attention. We observed the deposition of blue-fluorescing cuticular phenolics over guard and epidermal cells, as well as stomatal formation and patterning using the alkali-induced blue fluorescence of the cuticle of Prunus persica leaves. Stomata of different stages of maturity occurred together during leaf development, mainly at the tip of the lamina. The deposition of fluorescing compounds initially appeared over the guard cells of the developing stomata complexes and gradually extended to the neighbouring epidermal cells. Based on the blue fluorescence emitted by the cuticular layers, we constructed digital maps of leaves of different developmental stages, showing the pattern of stomatal formation and deposition of fluorescing compounds. A longitudinal tip-to-base gradient in the formation of stomata, as well as in the deposition of fluorescing compounds was observed in young developing leaves. The deposition of blue-fluorescing phenolic compounds seems to be coordinated with stomatal development.     

A systematic investigation of leaf epidermis in <Emphasis Type="Italic">Camellia</Emphasis> using light microscopy

The characters of mature leaf epidermis of 58 species and 3 varieties belonging to 19 sections, representing all the 4 subgenera: Protocamellia, Camellia, Thea and Metacamellia were investigated under light microscope. The main conclusions are as follows: 1) The shape of the epidermal cells is elliptical, polygonal or irregular and the anticlinal walls are arched, straight or sinuolate, sinuous to sinuate. 2) The stomatal apparatus, consisting of a stoma (a pore plus a pair of guard cells) and 3–4 subsidiary cells, restricted to the abaxial surface of the leaves, were assigned to anisocytic, anisotricytic, isotricytic or tetracytic type. 3) Special structures in leaf epidermis, such as stomatal clusters, aborted stomata, secretory cells, cuticular intrusions and their taxonomic implications were also elucidated.     

Guard cell wall: immunocytochemical detection of polysaccharide components

The composition of guard cell walls in sugar beet leaves (Beta vulgaris L.) was studied by using histochemical staining and immunocytochemical detection of cell wall antigens. The findings were compared with those in the walls of epidermal and mesophyll cells. Probing of leaf sections with monoclonal antibodies against pectins, terminal fucosyl residues linked alpha-(1-->2) to galactose, beta-(1-->3)-glucans and arabinogalactan-proteins revealed several specific features of guard cells. Pectic epitopes recognized by JIM7 were homogeneously distributed in the wall, whereas pectins recognized by JIM5 were not found in the walls themselves, but were abundant in the cuticular layer. Large amounts of molecules bearing terminal fucose were located predominantly in ventral and lateral guard cell walls. Much smaller amounts were detected in dorsal walls of these cells, as well as in the walls of pavement and mesophyll cells. Conspicuous accumulation of these compounds was observed in the vicinity of the guard cell plasmalemma, whereas labelling was scarce in the areas of the wall adjacent to the cell surface. The presence of callose clearly marked the ventral wall between the recently formed, very young guard cells. Callose also appeared in some mature walls, where it was seen as punctate deposits that probably reflected a specific physiological state of the guard cells. Large amounts of arabinogalactan-proteins were deposited within the cuticle, and smaller amounts of these proteoglycans were also detected in other tissues of the leaf. The histochemical and immunocytochemical structure of the guard cell wall is discussed in the light of its multiple functions, most of which involve changes in cell size and shape.     

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     

Observations on the cotyledonary and hypocotyledonary stomata and trichomes in some Caesalpiniaceae with a note on their taxonomic

The structure and ontogeny of stomata on cotyledons and hypocotyls and the trichomes on hypocotyl are accounted for in eighteen species of Caesalpiniaceae. Trichomes are eglandular, bi- rarely tri-celled, smooth walled or walls wavy with cuticular striations or tubercles. Anomocytic, haplocytic, paracytic, diacytic, transitional, tetracytic, tricytic and cyclocytic stomata occur in different combinations even on the same surface of the cotyledon. In all, there are fourteen combinations. Inspite of the diversity, the most frequent type is anomocytic in most of the species and paracytic in some species of Cassia and Delonix (abaxial) but rarely it is haplocytic or anisocytic. In hypocotyls it is anomocytic. Ontogenetically anomocytic, tetracytic and cyclocytic stomata are perigenous, whereas other types are mesogenous or mesoperigenous. There is an increase in the number of subsidiary cells by their division or the neighbouring perigenes assuming their shapes. About eight such types are described. A pair of stomata has a common subsidiary cell. Twelve types of guard cell and stomatal approximation abnormalities are described. A range in the number, size and shape of the nuclei in guard cell are recorded. Megastomata (giant stomata) are observed in Parkinsonia and Tamarindus. The taxonomic significance of the stomata is also discussed.     

黄瓜叶表面特性与生态适应性

对不同类型的黄瓜品种的气孔形态进行了扫描电镜观察 ,发现需水量不同的黄瓜品种气孔形态特征有很大的差异。保卫细胞表观差异比较明显 ,气孔的密度、大小及开张度均有显著的差异 :亚洲类型的黄瓜品种气孔密度较欧洲类型与欧亚杂交品种大 ,气孔的保卫细胞的大小为欧洲品种 >欧亚杂交品种 >亚洲品种。气孔周围的形态特征表现为 :亚洲品种保卫细胞角质褶皱较多 ,起伏较大 ,气孔深深地陷于褶皱中 ;欧洲品种黄瓜气孔保卫细胞周围角质则较平坦 ,保卫细胞突出于表皮细胞之上 ;欧亚杂交品种的气孔大小与形态介于上述二者之间。土壤含水量的不同也影响黄瓜叶片气孔的形态特性 ,随着土壤含水量的减少 ,黄瓜叶片单位面积的气孔密度增加 ,保卫细胞的大小变小 ,气孔开展度增加。     

Structure of Polymorphic Stomata in Canella winterena (L.) Geartn. (Canellaceae)

The presence of 10 types of normal stomata along with 4 subtypes, one intermediate type between stephanocytic and hemiparacytic and some abnormal stomatal structures both in the vegetative and floral organs of Canella winterena (L.) Geartn. (Canellaceae) is reported here. The epidermal cells are polygonal, isodiametric in surface view with straight anticlinal walls. The stomata are amphibrachyparacytic, anomocytic, anisocytic, brachyparacytic, brachyparatetracytic, cyclocytic, hemiparacytic, laterocytic, holoparacytic, stephanocytic, one intermediate type between stephanocytic and hemiparacytic, etc. Abnormalities like contiguous stomata, different types of twin stomata and cytoplasmic bridge between adjacent stomata, stomata with single guard cell, single guard cell with pore juxtaposed with normal stoma are also found. The presence of giant stomata is a significant finding. (© 2014 WILEY‐VCH Verlag GmbH & Co. KGaA, Weinheim)     

Epidermal studies in some Indian cultivars of Bougainvilleas

Epidermal studies in fifteen Indian cultivars of Bougainvilleas are described. The epidermal cells are polygonal isodiametric, or elongated with thick straight arched or slightly sinuous walls. Parallel culticular striations are radiating from guard cells. The mature stomata are anomocytic, paracytic and with a single subsidiary cell. The abnormal types noticed are: single guard cells with or without pores, arrested development, variously oriented contiguous stomata, cytoplasmic connections between nearby stomata and epidermal or subsidiary cells, and persistent stomatal cells. The development of anomocytic stomata is perigenous while that of the other types is mesogenous. Fifteen cultivars of Bougainvilleas are separated on the basis of bract colour, stomatal frequency and index per unit area.     

Epidermal structure and development of stomata in vegetative and floral organs ofHybanthus enneaspermus (Linn.)F. Muell

The present paper deals with the epidermal structure and ontogeny of stomata in vegetative and floral organs ofHybanthus enneaspermus. The epidermal cells are either polygonal or elongated with straight, sinuous or arched thick anticlinal walls. The surface of the cuticle shows parallel striations radiating from the guard cells or hair bases. Unicellular and uniseriate bicellular trichomes with verrucose margins have been observed on all organs. The mature stomata are anisocytic, paracytic, anomocytic and transitional between anisocytic and paracytic. The ontogeny of anisocytic and paracytic stomata is syndetocheilic or mesogenous, anomocytic is haplocheilic or perigenous, while that of the transitional type is mesoperigenous. Four types of stomata have been observed on all the vegetative and floral organs and their ontogeny from organ to organ of this plant is constant. Stoma with a single guard cell is the result of disintegration of one of the guard cells before or after pore formation. Contiguous stomata are also occasionally noticed.     

Developmental changes in guard cell wall structure and pectin composition in the moss Funaria: implications for function and evolution of stomata

Background and Aims

In seed plants, the ability of guard cell walls to move is imparted by pectins. Arabinan rhamnogalacturonan I (RG1) pectins confer flexibility while unesterified homogalacturonan (HG) pectins impart rigidity. Recognized as the first extant plants with stomata, mosses are key to understanding guard cell function and evolution. Moss stomata open and close for only a short period during capsule expansion. This study examines the ultrastructure and pectin composition of guard cell walls during development in Funaria hygrometrica and relates these features to the limited movement of stomata.

Methods

Developing stomata were examined and immunogold-labelled in transmission electron microscopy using monoclonal antibodies to five pectin epitopes: LM19 (unesterified HG), LM20 (esterified HG), LM5 (galactan RG1), LM6 (arabinan RG1) and LM13 (linear arabinan RG1). Labels for pectin type were quantitated and compared across walls and stages on replicated, independent samples.

Key Results

Walls were four times thinner before pore formation than in mature stomata. When stomata opened and closed, guard cell walls were thin and pectinaceous before the striated internal and thickest layer was deposited. Unesterified HG localized strongly in early layers but weakly in the thick internal layer. Labelling was weak for esterified HG, absent for galactan RG1 and strong for arabinan RG1. Linear arabinan RG1 is the only pectin that exclusively labelled guard cell walls. Pectin content decreased but the proportion of HG to arabinans changed only slightly.

Conclusions

This is the first study to demonstrate changes in pectin composition during stomatal development in any plant. Movement of Funaria stomata coincides with capsule expansion before layering of guard cell walls is complete. Changes in wall architecture coupled with a decrease in total pectin may be responsible for the inability of mature stomata to move. Specialization of guard cells in mosses involves the addition of linear arabinans.     

Foliar Penetration of Succinic Acid-2,2-dimethylhydrazide: Mechanism and Rate Limiting Step

The foliar penetration of succinic acid-2,2-dimethylhydrazide (SADH) into primary leaves of bean ( Phaseolus vulgaris L. cv. Blue Lake) was studied using detached leaves and a leaf disc method. SADH penetration was linear with time and directly related to concentration and the fraction of non-ionized SADH molecules in the treating solution. The addition of a surfactant, polyoxyethylene sorbitan monolaurate, increasing temperature and light enhanced penetration. Surfactant and light enhanced penetration are discussed in relation to increased wetting and area of contact over guard and accessory cells and the greater permeability of the cuticular ledges in open than in closed stomata. Evidence is presented that foliar penetration of SADH takes place by diffusion controlled by the cuticular membrane. Metabolic processes were not rate limiting.     

Studies on isolated starch-containing (Vicia faba) and starch-deficient (Allium cepa) guard cell protoplasts

Guard cell protoplasts from starch-containing Vicia faba and starch-deficient Allium cepa stomata were isolated, stabilized and recovered with an efficiency — in relation to the potential yield — of approx. 62% and 77%, respectively. In vitro, guard cell protoplasts (GCP) respond to abscisic acid and fusicoccin by respectively contracting and swelling, that is, decreasing or increasing in diameter by about 15% and more in comparison to the control. This in vitro response correlates with, but is more than 4 times as rapid as, the in vivo response of the stomata. Among the advantages presented by working with isolated GCPs are: greater sensitivity in response; freedom from influences of cuticular ridges, cell walls, subsidiary cells, and epidermal cells; and direct and parallel comparisons of starch-containing and starch-deficient GCP systems.Abbrecviations ABA abscisic acid - FC fusicoccin - ECP, MCP, and GCP epidermal, mesophyll, and guard cell protoplasts, respectively - PPV packed protoplast volume     

云南腾冲新近纪两种被子植物化石的角质层构造及其古环境意义

重点描述云南腾冲晚第三纪两种被子植物化石Betula mioluminifera Hu et Chaney,Carpinus subcordata Nathorst的角质层构造,并分析它们的现存最近亲缘种B.luminifera Winkler和C.cordata B1.var.mollis Cheng et Chen的表皮特征。实验分析证明：化石叶片的气孔参数可以推测地质历史时期大气CO2的浓度,并进而分析古环境的变化。C．subcordata Nathorst叶片能作为大气CO2浓度的生物指标。     

Organisation of microtubules and actin filaments in the cortex of differentiating Selaginella guard cells

Summary Using fluorescent probes and confocal laser scanning microscopy we have examined the organisation of the microtubule and actin components of the cytoskeleton in kidney-shaped guard cells of six species of Selaginella. The stomata of Selaginella exhibit novel cytoskeletal arrangements, and at different developmental stages, display similarities in microtubule organisation to the two major types of stomata: grass (dumbbell-shaped) and non-grass (kidney-shaped). Initially, cortical microtubules and F-actin radiate from the stomatal pore and extend across the external and internal periclinal cell surfaces of the guard cells. As the stomata differentiate, the cytoskeleton reorients only along the internal periclinal walls. Reorganisation is synchronous in guard cells of the same stoma. Microtubules on the inner periclinal walls of the guard cells now emanate from areas of the ventral wall on either side of the pore and form concentric circles around the pore. The rearrangement of F-actin is similar to that of microtubules although F-actin is less well organised. Radial arrays of both microtubules and F-actin are maintained adjacent to the external surfaces. Subsequently, in two of the six species of Selaginella examined, microtubules on both the internal and external walls become oriented longitudinally and exhibit no association with the ventral wall. In the other four species, microtubules adjacent to the internal walls revert to the initial radial alignment. These findings may have implications in the development and evolution of the stomatal complex.Abbreviations GC guard cell - MT microtubule     

Some new observations and interpretations on the vital staining of leaf epidermal tissue by neutral red

Summary The lower leaf epidermis from 5 plant species was stained with neutral red at 2 pH's (7.1 and 5.6) in the light and dark when the stomata were open or closed. At pH 5.6 no globule (= droplet) formation was observed in the guard cells whether the stomata were open or closed and cell walls possessed a high affinity for the stain. At pH 7.1 globules appeared in guard cells of open stomata, but not closed stomata, within 15 minutes. Anaerobic conditions prevented this globule formation. InZea mays, globules also appeared in subsidiary cells when the stomata were closed and in certain epidermal cells. Where globule formation did not occur increased diffuse staining of certain epidermal cells was considered to be the indication of cell integrity. In old leaf material very large numbers of dark blue globules appeared in epidermal cells ofCommelina diffusa, C. communis andSenecio odoris and this was associated with cell senescence.The staining characteristics were discussed in terms of cellular K⁺, Cl^–, tannin and flavonoid content and vacuolar pH.