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.     

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.     

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.     

Light and scanning electron microscopy-based foliar morpho-anatomical comparison of selected family Rosaceae members distributed in District Lahore,Punjab, Pakistan

In the present study morpho-anatomical characterization of selected Rosaceae members distributed in District Lahore was performed. Light and scanning electron microscopy was used for systematic characterization of the selected 19 species. Distinguished morpho-anatomical features such as size and shape of epidermal cells, size and type of stomatal cells, size and shape of trichomes, oil droplets, and silica bodies were contrasted. Results reported remarkable variations which could be taxonomically useful in identification of these members. Polygonal epidermal cells were observed in Eriyobotraya japonica, Potentilla bifurca, Potentilla supina, and Prunus amygdalus. However, Prunus cerasus possessed irregular-shaped epidermal cells that can be distinguished from hexagonal epidermal cells of Prunus persica. Similarly, stomatal type varied among some members. Paracytic or perisocytic stomata were observed in E. japonica whereas P. bifurca observed paracytic and anisocytic stomata. Lengths of guard cells were also of variable sizes. The average length of guard cells ranged from 53 (52–54) μm to 74 (73–75) μm in abaxial view. Potentilla supina had biggest, while Rosmarinus officinalis had tiny guard cells. Trichomes were tubular, stellate, cylindrical, ribbon-like, glandular, and nonglandular. Silica bodies in the present investigation were bilobed, rounded, and oval-shaped. It is inferred that diverse anatomical features proved to be valuable taxonomic tools that could be fruitfully helpful in identification of plants at specific as well as generic level.     

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.     

Microscopic investigations of some selected species of Papilionaceae through SEM and LM from Skardu valley,northern Pakistan

Pollen morphology of 10 species and foliar epidermal anatomy of eight species of Papilionaceae from Skardu valley, northern Pakistan has been estimated for the first time. The present study was commenced with an aim to provide a detailed account of the pollen morphology by scanning electron microscopy and foliar epidermal anatomy by light microscopy. The pollen aperture was tricolporate with reticulate exine in the selected species. Stomata types are actinocytic, paracytic, and anomocytic. Irregular or polygonal with undulate or straight walls, epidermal cells were reported. A unique diversity was observed in the foliar trichomes that show the taxonomic significance of the discrimination of taxa. Non‐glandular trichomes were observed in the selected species which are unicellular with thin, long and pointed apical cells. Pollen and foliar micro morphological characters proved to be helpful for the identification of taxa at a specific level.     

Scanning electron and light microscopy of foliar epidermal characters: A tool for plant taxonomists in the identification of grasses

SEM and light microscopy are of special interest for biologist to observe various features of the living bodies. In the current study we observed the foliar epidermal micro‐morphological characters of 44 grass species using SEM and Light microscopy to assess their taxonomic utility for taxonomists in the identification process. The aim of this study is to use the foliar epidermal structural variations in both upper and lower surfaces for identification of grasses. Significant diversity was observed in both qualitative and quantitative characters using SEM and Light microscopy. Variations were observed in stomatal number, size, guard cells shape, silica bodies, macro‐hairs, micro‐hairs, epidermal cell number, subsidiary cells, prickles, hooks, papillae, and short and long cells. A taxonomic key is prepared using these variations for the identification of grass species. Based on these findings, SEM and Light microscopy of foliar epidermal features can be of special interest for taxonomists in the identification of complex grass 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 morphology of leaf epidermis in eight populations of Atlas Pistachio (Pistacia atlantica Desf., Anacardiaceae)

A comparative analysis was undertaken to conduct a micromorphological study of Pistacia atlantica leaves by comparing different populations grown under different climatic conditions. Leaf epidermis of eight wild populations was investigated under scanning electron microscope. Micromorphological characteristics (epidermis ornament, stomata type, waxes as well as trichomes) of the adaxial and abaxial leaf surfaces were examined. The epidermis ornament varied among populations and leaf surface, the abaxial leaf surface is reticulate with a striate surface. Messaad site shows a smooth uneven surface. The adaxial leaf surface is smooth but several ornamentations can be seen. The leaflet is amphistomatic; the stomata appeared to be slightly sunken. A variety of stomatal types were recorded; actinocytic and anomocytic types are the most frequent. The indumentum consisted of glandular and nonglandular trichomes. Unicellular glandular trichomes are recorded for P. atlantica leaves in this study. Their density is higher in Oued safene site, located at the highest altitude in comparison with the other populations. The wax occurred in all the sites and its pattern varied according to the populations studied, particularly between Berriane and Messaad. The morphological variability exhibited by the eight populations of P. atlantica may be interpreted as relevant to the ecological plasticity and the physiological mechanisms involved are discussed in this report.     

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.     

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.     

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.     

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.     

Foliar micromorphology and its role in identification of the Apocynaceae taxa

In the present study, we evaluate the importance of foliar epidermal micromorphological characteristics of Apocyanaceae for accurate identification and classification. The species were collected from the University of Peshawar's main campus in the spring season to observe its qualitative and quantitative features. The length and width of guard cells, stomatal pore and subsidiary cells, trichomes, and crypts on both sides of the leaf were examined. Many species were observed to be hypostomatic. Plumeria rubra, Raulfia serpentine, Thevetia peruviana, Trachelospermum lucidum, Alstonia scholaris, and Catharanthus roseus demonstrated hypostomatic leaves. Nearly all the investigated species had anisocytic type of stomata only or in combination with other types of stomata on the upper and lower epidermis. Carissa carandas had anomocytic, anisocytic, and cyclocytic type of stomata on the upper epidermis, and the lower epidermis showed variations in stomatal type, such as anomocytic, stephanocytic, brachyparacytic, and hemiparacytic. Nerium oleander had no specific shape of stomata but showed stomatal crypts in which the stomata were enclosed inside many trichomes. The taxonomic key based on stomatal types, epidermal cells, stomatal index value, and statistical analysis, along with the variations in the epidermal cells, shows the link between the selected plants species, which will provide a baseline for future anatomical studies. This study highlights many undocumented micromorphological characteristics. The anatomical characteristics observed in this study will be helpful for taxonomic identification and species delimitation of the family Apocynaceae.     

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.     

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.     

Taxonomic study of subfamily Nepetoideae (Lamiaceae) by polynomorphological approach

The present study is insight into pollen morphology for characterizing species and their utility in the taxonomic separation of certain taxa of subfamily Nepetoideae (Lamiaceae) from Pakistan. The pollen micromorphology of 11 species of the Nepetoideae was analyzed and documented using light microscopy and scanning electron microscopy (SEM) for both qualitative and quantitative characteristics. Most species have hexazonocolpate pollen grains but trizonocolpate and tetrazonocolpate pollen with circular and oval amb were also rarely observed in Mentha spicata. The basic pollen shape in most of the studied species was subspheroidal but prolate grains were also observed in M. spicata, S. coccinea, and S. plebeia. The exine sculpturing of Nepetoideae pollen was taxonomically very informative particularly at subfamily level. Observations of exine sculpturing with SEM revealed various types of pollen grains: reticulate, bireticulate, microreticulate, perforate, aerolate, and gammate. The bireticulate type further subdivided into three subtypes based on the number of secondary lumina in each primary lumen and is characterized by varying characteristics of the secondary reticulum and primary muri. A significant variation was observed in colpus surface ornamentation. The maximum polar diameter was found in O. americanum (58 ± 5.8 μm) and the maximum equatorial diameter observed in O. basilicum (50.25 ± 1.37 μm). Pollen features of the studied species were discussed and compared based on the current taxonomical concepts. The results showed that pollen traits of the subfamily Nepetoideae was found significant to classify the taxa. Furthermore, pollen features provide additional evidence to distinguish macromorphologically similar taxa from each other.     

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.     

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.     

Seed morphology using SEM techniques for identification of useful grasses in Dera Ghazi Khan,Pakistan

In recent study, 15 taxa of family Poaceae from Dera Ghazi Khan were investigated for morphological characterization of seeds. Scanning electron microscopy (SEM) was utilized to study morphological features of grass seeds. Moreover, economic importance of studied taxa has been discussed in present research. Grass seeds or caryopsis were collected in their mature state from different areas of Dera Ghazi Khan. Collected caryopses were subjected to SEM to investigate surface sculpture, epidermal cell shape, and cell wall patterns. Caryopsis shape observed was elliptic to oval. Also, the morphology of plant is being investigated. Cell wall pattern observed was straight and wavy. Surface sculpturing varies from smooth to reticulate. Epidermal cell shape was either wavy or irregular. Hilum position is terminal in all species. Epidermal cell structure and cell wall pattern was not observable in some species. Major variations among studied taxa were observed in terms of seed surface pattern. Five types of caryopsis surface pattern were observed namely, striate, smooth, reticulate, papillate, and granulate. Investigated taxa illustrated variations in terms of studied morphological features. Utilization of SEM has been proved very helpful in exploring seed morphological features. These observed features can aid in delimitation and identification of various grass taxa. Recent research recommends the utilization of SEM for caryopsis morphology to solve the identification issue of problematic grass genera.