Foliar and stem epidermal anatomy of the tribe Cynoglosseae (Boraginaceae) and their taxonomic significance

Foliar and stem epidermal anatomical features of the tribe Cynoglosseae have been studied in detail for the taxonomic identification using light microscopy (LM) and scanning electron microscopic (SEM) techniques. A comparative study was conducted on different plant parts (leaf and stem epidermal anatomy) of 25 species belonging to eight genera of the tribe Cynoglosseae (Boraginaceae) collected from different phytogeographical regions of Iran for the first time. Different qualitative and quantitative characteristics were observed in detail using LM and SEM. Results showed that although generally the stem and leaf anatomical traits were similar, but some diagnostic features were examined for distinguishing the closely related genera in the tribe. The ratio of cortex/diameter of stem and phloem/xylem, the average row number of collenchyma, palisade and spongy cells, structure of trichomes, type of indumentum and palisade arrangement were found taxonomically important. The anatomical characters were statistically analyzed using cluster analysis and principal component analysis. The study found that stem and leaf eccentrics are variable in the genus but constant within species of the same genus. Most species had typical isobilateral leaves, but some showed an incipient dorsoventrally symmetry with a layer of abaxial palisade tissue. Eglandular trichomes were observed found in all the studied species, which were recognized based on structure and function. In present study some novel characters have been observed which are of great interest to the taxonomist for the correct identification some genera delimitations. The characters studied here are of less taxonomic value and delimitating at species level.     

Comparative anatomy on the vegetative organs of genus Ziziphora L. (Lamiaceae) from Turkey

The genus Ziziphora L. (Lamiaceae) is represented by five species (nine taxa) in the Turkish Flora. These taxa are Z. clinopodioides Lam. (subsp. elbursensis, subsp. filicaulis, subsp. kurdica, subsp. rigida), Z. capitata L., Z. persica Bunge, Z. tenuior L., Z. taurica Bieb. subsp. taurica, and Z. taurica Bieb. subsp. cleonioides (Boiss.) Davis which to be an endemic taxon for Turkey. They are strongly aromatic herbs which contain rich pulegone and used as herbal teas and spices and for this reason. In this study, comparative anatomy of the genus Ziziphora growing in Turkey is presented for the first time. In anatomical studies, cross sections of vegetative organs such as the root, stem, and leaf (lamina and petiole) were examined. In addition, to exhibit stomatal distribution and anatomy on adaxial and abaxial leaves were taken surface sections of the lamina and calculated stomatal index. Lamina and petiole anatomy were shown to be of great importance in the taxonomy of the Ziziphora taxa. The presence or absence of sclerenchyma in midrib of lamina and petiole, cortex parenchyma layer, mesophyll structure, and epidermal surface were found to be important characters for identification of Ziziphora taxa.     

Light microscopy of Pakistani Berberis leaf cuticles and its taxonomic implications

Taxonomy of the genus Berberis is quite complex, due to overlapping morphological characters, making it very difficult to differentiate the species within the genus. In order to resolve this taxonomic complexity, the foliar anatomy of 10 Berberis L. species was carried out, for the first time from Pakistan, using light microscopy (LM). Significant variation in terms of epidermal cells shape, size, cell wall pattern, and stomata type was observed. B. baluchistanica has the largest epidermal cells, Adaxial: length = 45–(53.9 ± 3.6)–62.5 μm; and width = 22.5–(26.3 ± 1.3)–30 μm; Abaxial: length = 37.5–(43.25 ± 2.5)–50 μm; and width = 20–(22.6 ± 0.8)–25. The highest number of stomata was observed in B. glaucocarpa as 62 on the abaxial surface while the lowest number of stomata was recorded in B. baluchistanica as 8 on the adaxial surface. Of 10 investigated species, 6 possess anomocytic type stomata, while 2 species that is, B. aitchisonii and B. parkeriana have both anomocytic and anisocytic stomata while B. baluchistanica and B. calliobotrys have only paracytic type stomata. The highest number of cells per unit area was present on the adaxial surface of B. calliobotrys ranging from 245–(252.4)–260 followed by B. parkeriana with 209–(227.8)–250 on the abaxial surface. Stomatal index (SI) also varied considerably and was the lowest (2.6) percentage in B. baluchistanica and highest (31.9) percentage in B. kunawurensis. A taxonomic key based on micro‐morphological characters is provided for species identification.     

Peltate trichomes on biogenic silvery leaves of Elaeagnus umbellata

External and internal features of Elaeagnus umbellata leaves were investigated by optical and electron microscopy. The adaxial (upper) and abaxial (lower) leaf surfaces appeared green and silver in color, respectively. There were peltate trichomes on both the adaxial and abaxial leaf surfaces. The peltate trichomes were 200–300 μm in diameter and shield or umbrella‐shaped. They had a central dome and 20–30 radiating rays that were fused to form a circular cap in the center and tapered at the end. The density of peltate trichomes was apparently higher on the abaxial leaf surface than on the adaxial leaf surface. At least two layers of peltate trichomes were commonly observed on the abaxial leaf surface. The epidermal cells on the abaxial leaf surface had convex lens‐like shape in cross sections. No distinct chloroplasts were found in the cytoplasm of peltate trichomes. These results suggest that the silver coloration on the abaxial leaf surface is mostly due to structural coloration associated with the profuse overlapping peltate trichomes having a circular cap of radially fused rays. The shrub did not show any pigmentary cellular features associated with the silver coloration. With the silvery leaves as a reflective surface for shaded leaves in canopy, E. umbellata is likely to adapt to the harsh non‐native light‐demanding environments.     

Comparative foliar anatomical and pollen morphological studies of Acanthaceae using light microscope and scanning electron microscope for effective microteaching in community

In this study, foliar anatomy and pollen morphology of 10 species of Acanthaceae has been investigated using light and scanning electron microscopy. The study was aimed to highlight the role of microscopy in microteaching at community for proper characterization of plants using palyno‐anatomical characters including pollen type, exine sculpturing, shape of epidermal cells, pattern of anticlinal wall, type and size of stomata, and trichome. Most of the species have polygonal cell shapes but some species have irregular, tetragonal, and pentagonal shape of epidermal cells. The largest epidermal cell length on adaxial and abaxial surface were observed in Asystasia gangetica 66.95 and 87.40 μm whereas least was observed on adaxial surface in Justicia adhatoda 36.9 μm and on abaxial surface in Barleria cristata 35.65 μm. In anatomy, species have diacytic type of stomata, whereas stomata of paracytic type observed in two species, while in A. gangetica cyclocytic type of stomata are present. Quantitively on abaxial surface, largest stomata length 29.9 μm and width 24.30 μm was noted in B. cristata. While shortest stomata length was observed in Ruellia prostrata 25.95 μm whereas minimum width of stomata was examined in Barleria acanthoides 2.05 μm. The diversity of trichomes are present in all species except in Ruellia brittoniana. Acanthaceae can be characterized by exhibiting different pollen morphology having five types of pollen shapes, prolate, spheroidal, perprolate, subprolate, and oblate spheroidal. Exine peculiarities showing variations such as reticulate, granulate, coarsely reticulate, lophoreticulate, perforate tectate, and granulate surface were examined.     

Palyno-anatomical characters and their systematic significance in the family Apiaceae from Chitral,eastern Hindu Kush,Pakistan

The present study was performed to provide a detailed explanation of leaf epidermal anatomy and pollen micromorphological features of selected species of family Apiaceae from Chitral, eastern Hindu Kush region as the basis of forthcoming studies. In the present article pollen morphology of eight species and foliar epidermal of seven species of family Apiaceae have been examined through microscopic techniques. In results two types of pollen prolate (five species) and perprolate (three species) with three colpi have been recorded. The exine ornamentation was found to be regulate, striate, and cerebroid. Largest pollen was found in Heracleum leucocarpum with the polar diameter of 43.25 μm and equatorial diameter of 21.6 μm. Smallest pollen was observed in Elaeosticta chitralica with the polar diameter of 18.4 μm. The P/E ratio varied from 1.59 to 2.16. Regarding to foliar epidermal anatomy, three types of epidermal cells including rectangular, irregular, and polygonal with variation in anticlinal wall pattern were determined. In the selected species three kinds of stomata comprising anisocytic, anomocytic, and paracytic type were reported in the current research. The size of epidermal cells ranged from 106 × 42.50 μm in Bupleurum falcatum subsp. cernuum and 77.25 × 26.35 μm in Prangos pabularia in adaxial surface. Largest stomatal complex was found in Prangos pabularia both in adaxial 33.55 × 20.05 μm and abaxial 50.25 × 39.40 μm. All the observed quantitative and qualitative features of the species were proved to be useful in the delimitation of species at generic and species level.     

Anatomy,micromorphology, and histochemistry of leaves and stems of Cantinoa althaeifolia (Lamiaceae)

This article enumerates the detailed anatomy of Cantinoa althaeifolia (Lamiaceae) illustrated with light and scanning electron microscopy images. The anatomical markers include the presence of branched nonglandular trichomes and capitate, peltate, and clavate types of glandular trichomes; prismatic crystals on the leaf and stem surfaces; and oil droplets in the leaf mesophyll. Histochemical tests and EDS analyses were performed in order to detect the composition of certain cells and their contents. The key findings of the present study can contribute to the taxonomy, species identification, and quality control of Cantinoa althaeifolia.     

Microscopic diagnosis of the leaf and stem of Piper solmsianum C.DC

Piper solmsianum C.DC., which is popularly known as pariparoba, is a shrub that measures 1–3 m in height and it inhabits areas with wet tropical soils. The objective of this study was to analyze the leaf and stem anatomy using light microscopy, scanning electron micrographs, and energy‐dispersive X‐ray spectroscopy in order to provide information for species identification. The anatomical profile showed the following main microscopic markers: hypostomatic leaf; hypodermis layer on both sides; pearl glands; biconvex midrib shape; five collateral vascular bundles in open arc with the central bundle larger than the others; circular stem shape; collateral vascular bundles arranged in two rings; sinuous sclerenchymatic sheath in the pith; secretory idioblasts; and starch grains in the mesophyll, in the ground parenchyma of the midrib, petiole, and in the stem; and six morphotypes of calcium oxalate crystals (styloids, cuneiform, tabular crystal rosettes, cuneiform crystal rosettes, elongated square dipyramids, as well as very elongated square dipyramids).     

Comparison of Bougainvillea spectabilis and Bougainvillea glabra species inhibited in Pakistan based on microscopic studies: Light microscope and scanning electron microscope

The anatomical variations of two plants from the Nyctaginaceae family, Bougainvillea spectabilis and Bougainvillea glabra, were studied using light and scanning electron microscopy methods in this work. Bougainvillea is a dicotyledonous with defensive traits that can withstand extreme (hot and dry) settings; according to the findings, crystal inclusions in cells, woody spines, and an abnormal development pattern are all features that help them survive against predators and are unique to this species. The Bougainvillea plant's leaves are arranged in simple pattern, alternate to each other along stem having an undulate leaves edge and an oval form. The xylem and phloem, palisade, parenchyma midrib, spongy mesophyll, raphide crystal bundles, and trichomes were all visible when bracts and leaves were transversally sectioned and dyed with toluidine blue O (TBO). The presence of crystals was confirmed by a detailed examination of the transverse leaves by using bright-field and cross-polarizing microscopy. Dissecting microscopic examination showed that all the leaves revealed leaves venation pattern that had midvein, lateral veins areoles, and trichomes. Although trichomes have been identified on both sides, a closer look at a cleaned leaf dyed with TBO showed multicellular abundant trichomes on adaxial surface. Stomata complexes were typically found on the abaxial surface of the leaf according to epidermal peels. Present studies also showed that on adaxial side, stomata were lesser in number or were absent and also showed that the morphologies of the pavement cells on the adaxial and abaxial sides of the leaf differed.     

Application of microscopy for discrimination of eight Swertia species utilized in the traditional Chinese medicine “Qingyedan”

Many Swertia species are utilized as a traditional medicine under the name “Qingyedan” in China, but are easily confused with one another. To distinguish eight Swertia species (S. mileensis, S. cincta, S. patens, S. punicea, S. delavayi, S. nervosa, S. macrosperma, and S. yunnanensis) and to ensure their safety and efficacy, the microscopic and macroscopic characteristics of the roots, stems, leaves, and flowers of them were examined. The results showed that microscopic and macroscopic features helpful for authentication of the eight species were the sinuosity of the anticlinal walls of epidermal cells and presence or absence of hairs on the leaf lamina; presence or absence of V‐shaped fibers and fibers with sinuous abaxial wall in the sepals; shape of epidermal cells and pattern of papillae on hairs on the margin of corolla nectary; distribution of stomata in leaf and sepal epidermises, stone cells in cortex and phloem of roots and in cortex and pith of stems, crystals in parenchymatous cells of mesophyll and stem, stomata size, stem diameter, and 4‐ or 5‐merous flowers, and so on. Two keys to the eight Swertia species based on macroscopic and microscopic characteristics are presented. The study indicates that microscopy and related techniques are convenient, practicable, and can be unambiguously applied for authentication of Swertia species. Microsc. Res. Tech. 76:296–310, 2013. © 2013 Wiley Periodicals, Inc.     

Electron microscopic observations of stomata,epicuticular waxes,and papillae in Chamaecyparis obtusa: Reconsidering the traditional concept of Y‐shaped white stomatal bands

The foliar morphological characters of hinoki (Chamaecyparis obtusa) were revisited using optical and scanning electron microscopy. In C. obtusa, typical Y‐shaped white stomatal bands were evident on the abaxial leaf surfaces. Two facial leaves and two lateral leaves were observed at the same node. Waxy papillae and oval stomata were arranged in two or three rows with protuberant rims on the abaxial leaf surfaces. Higher magnifications revealed the deposition of epicuticular waxes (tubules) on the Y‐shaped white stomatal bands. Given the absence of stomatal bands after dewaxing with organic solvents, the white stomatal bands in C. obtusa were related to the epicuticular waxes rather than the presence of aggregated stomata alone. In contrast to C. obtusa, a single median leaf and two lateral leaves were observed at the same node of oriental arborvitae (Platycladus koraiensis). Neither stomatal bands nor papillae were observed on P. koraiensis leaves. The stomatal density and epicuticular waxes in the stomatal regions of C. obtusa were higher than those of P. koraiensis. This study suggests that the traditional concept of Y‐shaped white stomatal bands in C. obtusa should be revised to describe the arrangement of the aggregated waxy stomata that occur in rows.     

Leaf epidermal micromorphology and its implications in systematics of certain taxa of the fern family Pteridaceae from Northern Pakistan

The present study is insights into foliar epidermal anatomy for characterizing clades, and their utility in taxonomic segregation of certain species of Pteridaceae from Northern Pakistan. The leaf epidermal anatomy of 10 species of Pteridaceae representing four genera were examined using light and scanning electron microscope. A micromorphological matrix was constructed for eight qualitative and 12 quantitative characters. unweighted pair group method with arithmetic means and principal components analysis statistical analysis were performed to test the validity of foliar epidermal anatomical features as method of separating species and genera, and phylogenetic clusters among species are constructed using qualitative and quantitative traits. The qualitative characters described here are shape of epidermal cells, stomata, guard cell and subsidiary cells, anticlinal wall pattern, and trichomes types which is helpful in defining groups within Pteridaceae. In addition, the size of stomata, guard cells, subsidiary cells, stomatal pore epidermal cells, and trichomes are quantitatively analyzed. All species have hypostomatic leaves. Two types of stomata were observed in studied species, anomocytic and polocytic. Anomocytic stomata were observed in three genera namely: Adiantum, Onychium, and Chielanthes whereas Pteris can be discriminated from other genera by its polocytic stomata. On the basis of multivariate analysis present study does provides sufficient information on the taxonomic importance of foliar anatomy which validate its efficacy in species and genera discrimination. From result obtained here it is further possible to use leaf micromorphologic data in ferns phylogeny and providing basis for future taxonomic delimitation in other taxa.     

Tillandsia stricta Sol (Bromeliaceae) leaves as monitors of airborne particulate matter—A comparative SEM methods evaluation: Unveiling an accurate and odd HP‐SEM method

Airborne particulate matter (PM) has been included among the most important air pollutants by governmental environment agencies and academy researchers. The use of terrestrial plants for monitoring PM has been widely accepted, particularly when it is coupled with SEM/EDS. Herein, Tillandsia stricta leaves were used as monitors of PM, focusing on a comparative evaluation of Environmental SEM (ESEM) and High‐Pressure SEM (HPSEM). In addition, specimens air‐dried at formaldehyde atmosphere (AD/FA) were introduced as an SEM procedure. Hydrated specimen observation by ESEM was the best way to get information from T. stricta leaves. If any artifacts were introduced by AD/FA, they were indiscernible from those caused by CPD. Leaf anatomy was always well preserved. PM density was determined on adaxial and abaxial leaf epidermis for each of the SEM proceedings. When compared with ESEM, particle extraction varied from 0 to 20% in air‐dried leaves while 23–78% of particles deposited on leaves surfaces were extracted by CPD procedures. ESEM was obviously the best choice over other methods but morphological artifacts increased in function of operation time while HPSEM operation time was without limit. AD/FA avoided the shrinkage observed in the air‐dried leaves and particle extraction was low when compared with CPD. Structural and particle density results suggest AD/FA as an important methodological approach to air pollution biomonitoring that can be widely used in all electron microscopy labs. Otherwise, previous PM assessments using terrestrial plants as biomonitors and performed by conventional SEM could have underestimated airborne particulate matter concentration.     

Leaf epidermal characteristics of Cissampelos L. (Menispermaceae) species from Northeastern Brazil

The morphological similarities among the species of Cissampelos are remarkable and the difficult to distinguish them as well. This article presents a comparative anatomical study of the leaves of common Northeastern Brazilian species of Cissampelos, carried out using light and scanning electron microscopy. The leaf epidermal was studied to obtain data on epidermal characteristics and to evaluate their taxonomic significance. As results, some micromorphological characters on the leaf epidermal like the cuticular waxes, the presence of papillae in epidermis and nonglandular trichomes, the anticlinal walls epidermal cells, the distribution, density and type of trichomes, and also the type and distribution of epicuticular wax proved to be the most useful characteristics to distinguish the species in taxonomic studies. Microsc. Res. Tech., 2011. © 2010 Wiley‐Liss, Inc.     

Morphoanatomical and phytochemical studies of Salvia lachnostachys (Lamiaceae)

Salvia lachnostachys Benth., Lamiaceae, is a endemic species from southern Brazil. The essential oil of its leaves and flowers is mainly constituted by aliphatic compounds, such as dodecanoic acid, with sesquiterpenes as minor constituents. This work evaluated the morphology, anatomy, microchemistry, and phytochemistry of S. lachnostachys to provide advanced knowledge of Brazilian plants with medicinal potential. Light and scanning electron microscopy techniques were used in the anatomical and microchemical studies. Compounds were isolated by chromatographic techniques, identified by analysis of their NMR spectra and compared with published data. S. lachnostachys can be distinguished from other related species mainly by its petiolate leaves, terminal inflorescence, persistent bracts, and villous‐glandular corolla. The stem and leaves of S. lachnostachys display anatomical characteristics common to the family Lamiaceae. However, this species can be distinguished from other family members by the morphology and the presence of eglandular and glandular trichomes, as well as the organization of the vascular bundles of the petiole. The phytochemical results revealed that S. lacnostachys produces oleanolic and ursolic acids in addition to the diterpene fruticuline A, which is a rare compound, previously found only in Salvia fruticulosa Benth. and S. corrugata Vahl. Ursolic and oleanolic acids are bioactive triterpenes that exhibit antiatherosclerotic, anticancer, antihypertensive, antinflammatory, antileukemic, antimutagenic, antioxidant, antiproliferative, and antiviral activities, and fruticuline A has antibacterial activity. Microsc. Res. Tech. © 2012 Wiley Periodicals, Inc.     

Leaf blade anatomy and ultrastructure of six <i>Simira</i> species (Rubiaceae) from the Atlantic Rain Forest,Brazil

Simira is a predominantly woody Neotropical genus comprising 41 taxa, 16 of which occur in Brazil and eight of them in the southeastern region of Brazil. Leaf blades of Simira eliezeriana Peixoto, S. glaziovii (K. Schum.) Steyerm., S. grazielae Peixoto, S. pikia (K. Schum.) Steyerm., S. rubra (Mart.) Steyerm., S. sampaioana (Standl.) Steyerm. were collected in the southeastern region of Brazil and fixed according to usual methods for light and electron microscopy. The leaf blades show typical characteristics of the Rubiaceae family as dorsiventral mesophyll and paracytic stomata. The presence of two bundle sheaths that extend to the upper epidermal layer, prismatic crystal and crystal-sand, alkaloids in the mesophyll and the organization micromorphological of the outer periclinal wall are considered characteristics representative for the genus. This study also demonstrates some leaf blade characteristics that can be used to Simira species identification (leaf surface, domatia types, epicuticular wax types and patterns of epidermis anticlinal cell walls).     

Systematics study through scanning electron microscopy; a tool for the authentication of herbal drug Mentha suaveolens Ehrh

In all over the world, herbal drugs are usually adulterated with similar species or varieties due to incorrect identification. Most of herbal products devoid purity and quality, therefore an attempt was carried out to identify plant species and authenticate its herbal drug products from Mentha suaveolens. Microscopy tools provide an excellent platform to identify plants at species level. In this study, microscopic and pharmacokinetic parameters of M. suaveolens were observed. Plant species were collected from high diverse areas of Northern Pakistan. Macro and micro‐morphology including palynology and anatomical features were analyzed to study M. suaveolens. Species characteristics were studied, while implementing microscopic techniques for the delimitation and identification of the species. Traditionally Mentha species are used to cure several diseases that is, digestive disorders, respiratory disorders. Micromorphology (stem, leaves, flowers structure, length etc.), palynology (shape, size of pollen etc.), and anatomical characters (types of stomata, epidermal cell shape, and trichomes) were studied. Micromorphology and anatomical characters were of great interest and significance to discuss the taxonomy of the species. Taxonomic characters were studied to characterize and authenticate the species. The aim of the present study is to observe in detail the taxonomic identification of the species in term of morphology, palynology, and foliar epidermal anatomy for the correct identification along with their medicinal uses in the area.     

Comparative morphology of the leaf epidermis in Lobelia (Lobelioideae) from China

The leaf epidermal morphology of 38 samples, representing 22 Lobelia species from China, was studied using light microscopy and scanning electron microscopy. The stomata and other epidermal features were largely consistent within species, and therefore represented good characters for taxonomic purposes. Diagnostic characters of the leaf epidermis were used to differentiate species of Lobelia. The species pairs L. clavata and L. pyramidalis, L. alsinoides and L. terminalis, and L. davidii and L. erectiuscula, were differentiated using leaf epidermal characters. Results at the micro‐structural level partly supported the current subdivisions of Lobelia. Previous taxonomic revisions of Lobelia in China were evaluated based on species epidermal characteristics. The results supported the recognition of Pratia merged with Lobelia, and of L. chevalieri as a distinct species. Lobelia brevisepala should not be included in L. montana. The status of Lobelia in the evolutionary history of Campanulaceae was assessed. The Lobelia species investigated retained many primitive leaf epidermal features.