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.     

Utilization of foliar cuticle morphology for the identification of weedy grasses

For the robust identification of weeds of taxonomically complex families like Poaceae, scanning electron microscopy (SEM) and light microscopy (LM) are of significant implications. In this study, SEM and LM methods were used by investigating 12 Pakistani weedy grass species as an example for foliar epidermal micromorphological characters for its role in the identification of grasses species. Qualitative and quantitative foliar micromorphological variations in stomatal type, number, guard cells, subsidiary cells, silica bodies and trichomes were studied. Significant variation was observed in both qualitative and quantitative features and based on this diversity in epidermal characters, a key was drawn to delimitate the species. Variations in stomata, silica bodies (SiO₂·nH₂O), macrohairs, microhairs, hooks, papillae, prickles, and other epidermal structures were used in establishing the taxonomic key. The data were also statistically evaluated by determining its mean, standard deviation, variance, the coefficient of variance, and standard error. Foliar epidermal characters such as silica bodies shape and trichomes were taxonomically more useful than stomatal types, subsidiary, and guard cells. SEM and LM micromorphological characters were of substantial importance in the delimitation of closely related species.     

Comparative SEM and LM foliar epidermal and palyno‐morphological studies of Amaranthaceae and its taxonomic implications

Palynological features as well as comparative foliar epidermal using light and scanning electron microscope (SEM) of 17 species (10genera) of Amaranthaceae have been studied for its taxonomic significance. Different foliar and palynological micro‐morphological characters were examined to explain their value in resolving the difficulty in identification. All species were amphistomatic but stomata on abaxial surface were more abundant. Taxonomically significant epidermal character including stomata type, trichomes (unicellular, multicellular, and capitate) and epidermal cells shapes (polygonal and irregular) were also observed. Pollens of this family are Polypantoporate, pores large, spheroidal, mesoporous region is sparsely to scabrate, densely psilate, and spinulose. All these characters can be active at species level for identification purpose. This study indicates that at different taxonomic levels, LM and SEM pollen and epidermal morphology is explanatory and significant to identify species and genera.     

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.     

Using microscopic techniques for taxonomic implications of seed and fruits of Delphinium L. (sensu lato) (Ranunculaceae)

In present research, micromorphological characters of the genus Delphinium L. (sensu lato) Ranunculaceae seeds and fruits were studied using microscopic techniques. A total of 37 species were studied using light (LM) and scanning electron microscopy (SEM) to evaluate seeds and fruits micromorphological characteristics for the taxonomic identification and species relationship in the genus. For each taxon, 33 characters were observed and studied. All characters were analyzed using MVSP software and UPGMA method by using dissimilarity index (average taxonomic distance). Considerable differences were observed in trichomes shape of fruit and seed color features. The results revealed that all studied taxa are separated into three clades. The first clade consists of D. pallidiflorum and D. semibarbatum, which have black seed and two types of glandular and crispate trichomes in fruit. The second clade consists of D. speciosum, D. szowitsianum, D. dasystachyson, D. aquilegifolium, D. ursinum, C. trigonelloides, and C. camptocarpa, which had a brown color seeds, two types of glandular and villous trichomes on fruits also have been observed. The remaining species were classified as third clade having various colors including yellow, black, and brown color seeds, the trichome shapes included one of the following forms strigose, villous, trichomes less, glandular, hooked, and crispate trichomes in fruits. The aim of the present study is to use micromorphological characters of seeds and fruits for the taxonomic identification and species delimitation in the genus Delphinium.     

Taxonomic significance of foliar epidermal morphology in Lamiaceae from Pakistan

Foliar micromorphological features are useful to elucidate the taxonomy and systematics of the Lamiaceae species. Leaf epidermal morphology using scanning electron microscopy and light microscopy of 22 Lamiaceae species from 15 genera have been investigated with an aim to solve its taxonomic problem in the correct identification. Various foliar micromorphological features were observed to explain their importance in resolving the correct identification of Lamiaceae taxa. Two main types of trichomes were observed; glandular trichomes (GTs) and nonglandular trichomes (NGTs). GTs were further divided into seven subtypes including the capitate, subsessile capitate, sessile capitate, sunken, barrel, peltate, and clavate. Similarly, NGTs were also divided into simple unicellular and multicellular including conical, falcate, cylindrical, dendrite, papillose, and short hook shape. Quantitative measurement includes the length and width of the trichomes, stomatal complex, epidermal cells, stomata, and trichomes index. Based on the foliar micromorphological characters, a taxonomic key was developed to delimit and correctly identify studied taxa. Further molecular, other anatomical and phylogenetic studies are recommended to strengthen the systematics of Lamiaceae.     

Foliar epidermal anatomy of Lamiaceae with special emphasis on their trichomes diversity using scanning electron microscopy

Foliar epidermal features were based on the micromorphology of trichomes types, epidermal cells and stomatal complex. Even though each feature has its own limited taxonomic value but collectively these characteristics may be systematically important especially for the discrimination and identification of complex and problematic taxa. The systematics significance of nonglandular (NGTs) and glandular trichomes (GTs), stomatal complex and epidermal cells of Lamiaceous flora were analyzed by using the light microscopy (LM) and scanning electron microscopy (SEM). Variations on the observed epidermal appendages were divided into two basic types: glandular and nonglandular. GTs can be divided into subtypes: sessile capitate, subsessile capitate, and barrel and sunken. NGTs were also divided into subtypes: dendritic, stellate, conical, falcate, simple and 1–6 cells long having granulate and smooth surface ornamentation. NGTs were the most dominant features of both adaxial and abaxial surfaces of all observed taxa. Vitex negundo, Isodon rugosus, Colebrookea oppositifolia, and Marrubium vulgare could be demarked because of their twisted like appearance of NGTs at the abaxial surface. The Lamiaceae had both hypostomatic and amphistomatic leaf. Stomata were observed as diacytic, anisocytic, and anomocytic. Epidermal cells were found to be irregular, isodiametric, and rectangular. Based on these characters a taxonomic key was developed to delimit the closely related taxa. Distribution and morphology of the foliar epidermal trichomes through SEM highlight an important taxonomic tool used by the taxonomists as an aid to the correct identification of problematic Lamiaceae taxa.     

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.     

Systematic approach to the correct identification of Asplenium dalhousiae (Aspleniaceae) with their medicinal uses

In present study, multiple microscope techniques were used for the systematics identification of the species Asplenium dalhousiae. The plant was collected from different phytogeographical and its natural habitat of Pakistan, where it shows higher diversity. Morphology, foliar epidermal anatomy, and spore morphological characters of the species were studied in detailed using multiple microscopic techniques through light microscopy (LM) and scanning electron microscopy (SEM). LM and SEM were used for the systematics identification of the species. Traditionally, the species is used in the ailment of many diseases, so the spore morphology, anatomical features, and morphological characters are relevant to describe the species taxonomy. The importance of multiple methods of taxonomic study (e.g., documentation and morphological characteristics) for characterizing herbs are important step in systematic certification to maintain the efficacy of herbal medicines. The aim of the present study is to examine the morphological, anatomical, and spore morphology of the species A. dalhousiae in more detailed for the correct taxonomic identification and their medicinal validation from Pakistan.     

Leaf epidermal characters of Solanum sect. polytrichum (Solanaceae) as taxonomic evidence

The morphological similarities among the species of Solanum are remarkable, and are often very difficult one clear distinction between them. This paper presents a comparative anatomical study of the leaf epidermis of five Brazilian species of Solanum sect. Polytrichum, carried out using light and scanning electron microscopy. The leaf epidermis surfaces were investigated to evaluate their taxonomic significance to be used for separation and delimitation of the species of the section. As results, some micro-morphological characters of the leaf epidermis, such as density, distribution and type of stellate trichomes, and the anticlinal walls of epidermal cells, and also the type and distribution of stomata proved to be the most useful and distinctive characters for the separation and delimitation of the species, and also may contribute as an additional support to the interspecific taxonomy and systematic of Solanum sect. Polytrichum.     

Comparative pollen and foliar micromorphological studies using light microscopy and scanning electron microscopy of some selected species of Lamiaceae from Alpine Zone of Deosai Plateau,Western Himalayas

The study was conducted to highlight a detailed account of morphology of pollen chosen species of Lamiaceae through scanning electron microscopy, and the anatomical characteristics of leaf epidermis of seven species using simple light microscopy. In results, Anisomeles indica and Otostegia aucheri belong to subfamily Lamioideae because it has tricolpate pollen while the rest eight species belong to subfamily Nepetoideae (hexacolpate pollen). The exine sculpturing of pollen of studied species was found to be reticulate. In the family Lamiaceae, four kinds of stomata were found anomocytic, anisocytic, diacytic, and actinocytic, respectively. The cell wall patterns of epidermal cells were irregular or polygonal with straight or undulate walls. It was noted that the variety of the epidermal trichomes seems of taxonomically important for the identification of species of Lamiaceae. Both nonglandular and glandular trichomes were analyzed. The nonglandular trichomes were characterized with long, thin, and pointed apical unicellular cells. The nonglandular trichomes were A‐shaped in Thymus linearis. In Perovskia abrotanoides, stellate glandular trichomes were observed whereas in A. indica and Mentha royleana both glandular and nonglandular trichomes were found. In A. indica, the nonglandular trichomes were sessile and peltate in M. royleana. For the first time in this study, pollen and foliar micromorphological features of selected species of this area are carried out. These taxonomic characters were found to be important in discrimination of species from each other. In future, the detailed study with comprehensive morphology coupled with other important characters is required for delimitation of taxa at various levels.     

Identification of monocot flora using pollen features through scanning electron microscopy

Pollen used to track structural and functional evolution in plants as well as to investigate the problems relative to plant classification. Pollen characters including ornamentation, shape, apertural pattern, pollen symmetry, colpus length, width, and margins used to detect the similarities and dissimilarities between genera and also species of the same genus. In this study pollen features of 20 monocot species belonging to 15 genera of the Amaryllidaceae, Asparagaceae, Iridaceae, Ixioliriaceae, Liliaceae, and Xanthorrhoeaceae were studied using scanning electron microscopy (SEM) and light microscopy (LM). In this study two species that is Zephyranthes citrina and Tulbaghia violacea were reported for the first time from Pakistan. Pollen grains were visualized with LM. Non‐acetolyzed and acetolyzed pollen were examined using SEM. A taxonomic key was developed to highlight the variation in pollen features in order to make their systematic application for correct species identification.     

Palyno‐anatomical studies of monocot taxa and its taxonomic implications using light and scanning electron microscopy

Palyno‐anatomical study of monocots taxa using Light and Scanning Electron Microscopy (SEM) was first time conducted with a view to evaluating their taxonomic significance. Studied plants were collected from different eco‐climatic zones of Pakistan ranges from tropical, sub‐tropical, and moist habitats. The aim of this study is to use palyno‐anatomical features for the correct identification, systematic comparison, and investigation to elucidate the taxonomic significance of these features, which are useful to taxonomists for identifying monocot taxa. A signification variation was observed in quantitative and qualitative characters by using the standard protocol of light microscopy (LM) and SEM. Epidermal cell length varied from maximum in Allium griffthianum (480 ± 35.9) μm at the adaxial surface to minimum in Canna indica (33.6 ± 8.53) μm on abaxial surface. Maximum exine thickness was observed in Canna indica (4.46) μm and minimum in Allium grifthianum (0.8) μm. Variation was observed in shape and exine ornamentation of the pollen, shape of the epidermal cell, number, size, and type of stomata, guard cell shape, and anticlinal wall pattern. Based on these palyno‐anatomical features a taxonomic key was developed, which help in the discrimination of studied taxa. In conclusion, LM and SEM pollen and epidermal morphology is explanatory, significant, and can be of special interest for the plant taxonomist in the correct identification of monocots taxa.     

Seed micromorphology and its taxonomic evidence in subfamily Alsinoideae (Caryophyllaceae)

Seed micromorphology of 13 species, belonging to four genera of subfamily Alsinoideae (Caryophyllaceae) were investigated with scanning electron microscopy (SEM), in order to assess their diagnostic significance at generic level and provide additional evidence on species delimitation, as well as correct identification and phylogenetic position. Genera and species of subfamily Alsinoideae exhibit great variation in ultrastructure and a high diversity of novel micromorphological characters were observed. Variation in seed shape, color, hilum, anticlinal wall, epidermal cell, cell surface, margins, and quantitative characters as length and width were studied in detail, compared, illustrated, and their taxonomic significant were discussed. Seed shapes of the species were classified as reniform, round, angular, subcircular, subreniform, and elliptical pyriform, with sub‐central, central, basal, and nearly basal hilum. Wavy, irregular, tetragonal, and elongated epidermal cells structure has been observed as an exomorphological character. The present findings show that the micromorphology of subfamily Alsinoideae provides taxonomic information and is helpful to distinguish different species. The results also explained that SEM morphology of seeds provide important data about affinity among taxa and give potential characters in delimitation of members of subfamily Alsinoideae at generic and species level. A principal component analysis allowed to highlight the most outsiders among seed micromorphology with a possible explanation. Taxonomic keys were developed based on micromorphological characters to delimit the species and useful for their quick identification within subfamily Alsinoideae.     

Intraspecific variation in seed morphology of tribe vicieae (Papilionoidae) using scanning electron microscopy techniques

Seed micromorphology of 12 species of tribe Vicieae (Papilionoidae) representing five genera were examined using Scanning Electron Microscope (SEM). The different seed types were described, illustrated, compared, and their taxonomic importance is discussed. Seeds exhibit great diversity in ultrastructure and a variety of novel morphological features have been determined among and within genera of the tribe. Seeds were characterized by oval to round shape, ovate to oblong outline and striate to papillate ornamentation. Radiate and wavy cell pattern has been observed as a characteristic exomorphological feature. In Vicia sativa, testa cells superficially seem to form peltate hairs. As a result, seed micromorphology with few exceptions showed variation and its taxonomic value was significant in distinguishing taxa at species level. The disparity in shape and density of papillae or protuberances and alignment of testa cells, may possibly give further insight at intraspecific level within tribe. The findings also illustrated that the use of SEM in seed morphology can reveal hidden morphological affinities among species and showed potential in delimitation of Vicieae members generally at tribe, genus, and particularly at species level.     

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.     

Morphological characterization of Hypnaceae (Bryopsida,Hypnales): Investigating four genera from Western Himalayas by using LM and SEM techniques

The Hypnaceae is one of the largest and diversified family among the pleurocarpous mosses which consists of nearly 60 genera and 1,000 species around the world. In Pakistan, it is represented by 15 species and 8 genera. The current research article provides the detail morphological investigation of four different species (genera) of the family Hypnaceae that is, Gollania clarescens, Hypnum revolutum, Homomallium andoi, and Taxiphyllum taxirameum in the Western Himalayas, Pakistan. The research article reports the first SEM study of the leaf surfaces of the studied taxa, along with comprehensive morphological characters of the four species. The main objective of the research project is to present the comparative light and scanning electron microscopic study to discuss the morphology in detail because previously the family is just reported in different bryophyte checklists of the Western Himalayas. Based on results, morphological characters, micromorphological observations, qualitative and quantitative attributes, taxonomic keys for the studied taxa, distribution, and habitat are described. Stereoscope was used to analyze different qualitative characters, and light microscope was used to observe and measure the laminal and alar cells of the leaf. The SEM study reveals many important surface features like cell shape and cell wall. The laminal cells were mostly linear and elongated with thin cell walls. The specimens used for the SEM were air dried, so the laminal cells were somewhat constricted and concaved. The current study project will help to make the contribution in the taxonomy and morphology of this family.     

Leaf and stem micromorphology of Byrsonima sericea DC. by light and scanning electron microscopy

Micromorphological studies were carried out using multiple microscopic techniques on the leaves and stem bark of Byrsonima sericea DC. (Malpighiaceae), a species popularly known as “murici” and used medicinally, in order to identify both qualitative and quantitative features of leaf and stem anatomy and histochemistry as differential parameters to support both the quality control of its ethnodrugs and the taxonomy of the genus. The study was conducted using traditional techniques of plant anatomy, histochemical tests, and the stomatal index (SI). Byrsonima sericea has hypostomatic leaves, anomocytic stomata, and its epidermal walls are anticlinal and straight on the adaxial and curved on the abaxial faces. T‐shaped trichomes were observed mainly on the abaxial surface. The leaf epidermis showed waxes syntopism on both surfaces, with the occurrence of different crystalloid forms on a single phylloplane. The mesophyll is dorsiventral, with 3‐4 collateral vascular bundles. Phenolic compounds, starch, and proteins were identified in the petiole and stem. The SI was 14.5 ± 0.53% (p < .05), but did not showed significant variations. A set of characters were found to be distinctive for the studied species, however, constituting parameters that could be used to separate B. sericea from other species of the genus.