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.     

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.     

An X-ray microscopy perspective on the effect of glutaraldehyde fixation on cells

X-ray microscopy (XRM) is the only microscopy technique that can provide high-resolution (30 nm) imaging of biological specimens without the need to fix, stain or section them. We aim to determine the effect, if any, of glutaraldehyde fixation on algae cells from the XRM perspective and thus provide beneficial information for both X-ray and electron microscopists on artefacts induced by glutaraldehyde fixation. Three species of microalgae, Microcystis aeruginosa, Anabaena spiroides and Chlorella vulgaris, were used in this study. XRM images were obtained from unfixed and glutaraldehyde-fixed cells and cell diameter and percentage X-ray absorbency were measured. The mean diameter of cells from fixed preparations was smaller than from unfixed preparations; the mean diameter of M. aeruginosa cells was significantly reduced from 3.92 µm in unfixed cells to 3.43 µm in fixed cells (P < 0.05); in C. vulgaris the diameter of cells was also significantly reduced from 3.50 µm in unfixed to 2.98 µm in fixed samples (P < 0.05); whereas there was no significant reduction in the diameter of A. spiroides cells (4.04–3.90 µm). The protein crosslinking mechanism of glutaraldehyde probably generated free water molecules, which play an important role in radiation damage induced by X-rays. This was seen as mass loss and cell shrinkage, which in the present study occurred more frequently in fixed cells than in unfixed cells. In addition, we demonstrated that the uptake of glutaraldehyde by cells makes all protein constituents in the cell organize into a closely packed configuration, thus causing a rise in the percentage of X-ray absorbency. In fixed cells, this rise was approximately by a factor of two compared with unfixed samples in which protein constituents inside the cell are arranged in their native form.     

Micro‐CT measurements of tumoral vessels supplied by portal circulation in hepatic colorectal metastasis mouse model

The purpose of this study was to elucidate the micro CT findings of tumoral vessels supplied by portal circulation during establishment of hepatic metastasis of colorectal cancer in a mouse model. Hepatic metastases were induced in 15 BALB/c mice through the injection of murine colonic adenocarcinoma tumor cells into the mesenteric vein. Micro‐CT imaging of the tumoral vessels was obtained to clarify the microvascular architecture. We evaluated the sinusoidal structure, diameter of the tumoral vessels (DTV) and blood vessel density (BVD) according to tumor sizes ranging from 201 to 3,000 µm in diameter. A total of 116 tumors were observed on day 15 after cell injection. The mean diameter of a normal hepatic sinusoid was 11.7 ± 2.0 µm on micro CT. The DTV supplied by the portal vein of tumors measuring 1,001–1,500 µm in diameter was greater than that of tumors 200–1,000 µm in diameter. The mean BVD from the portal vein gradually decrease according to size of tumor from 201 to 3,000 µm in diameter (r² = ?0.584, P < 0.01). The characteristics of tumoral vessels supplied by portal circulation during establishment of hepatic colorectal metastases were well visualized with micro‐CT imaging. Microsc. Res. Tech. 77:415–421, 2014. © 2014 Wiley Periodicals, Inc.     

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.     

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.     

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.     

Antimicrobial effect,frictional resistance,and surface roughness of stainless steel orthodontic brackets coated with nanofilms of silver and titanium oxide: a preliminary study

Nano‐silver and nano‐titanium oxide films can be coated over brackets in order to reduce bacterial aggregation and friction. However, their antimicrobial efficacy, surface roughness, and frictional resistance are not assessed before. Fifty‐five stainless‐steel brackets were divided into 5 groups of 11 brackets each: uncoated brackets, brackets coated with 60 µm silver, 100 µm silver, 60 µm titanium, and 100 µm titanium. Coating was performed using physical vapor deposition method. For friction test, three brackets from each group were randomly selected and tested. For scanning electron microscopy and atomic‐force microscopy assessments, one and one brackets were selected from each group. For antibacterial assessment, six brackets were selected from each group. Of them, three were immediately subjected to direct contact with S. mutans. Colonies were counted 3, 6, 24, and 48 h of contact. The other three were stored in water for 3 months. Then were subjected to a similar direct contact test. Results pertaining to both subgroups were combined. Groups were compared statistically. Mean (SD) friction values of the groups 'control, silver‐60, silver‐100, titanium‐60, and titanium‐100' were 0.55 ± 0.14, 0.77 ± 0.08, 0.82 ± 0.11, 1.52 ± 0.24, and 1.57 ± 0.41 N, respectively (p = .0004, Kruskal–Wallis). Titanium frictions were significantly greater than control (p < .05), but silver groups were not (p > .05, Dunn). In the uncoated group, colony count increased exponentially within 48 h. The coated groups showed significant reductions in colony count (p < .05, two‐way‐repeated‐measures ANOVA). In conclusions, all four explained coatings reduce surface roughness and bacterial growth. Nano‐titanium films are not suitable for friction reduction. Nano‐silver results were not conclusive and need future larger studies.     

Topographic characterization of unworn contact lenses assessed by atomic force microscopy and wavelet transform

This paper analyses the three‐dimensional (3‐D) surface morphology of optic surface of unworn contact lenses (CLs) using atomic force microscopy (AFM) and wavelet transform. Refractive powers of all lens samples were 2.50 diopters. Topographic images were acquired in contact mode in air‐conditioned medium (35% RH, 23°C). Topographic measurements were taken over a 5 µm × 5 µm area with 512 pixel resolution. Resonance frequency of the tip was 65 kHz. The 3‐D surface morphology of CL unworn samples revealed (3‐D) micro‐textured surfaces that can be analyzed using (AFM) and wavelet transform. AFM and wavelet transform are accurate and sensitive tools that may assist CL manufacturers in developing CLs with optimal surface characteristics. Microsc. Res. Tech. 78:1026–1031, 2015. © 2015 Wiley Periodicals, Inc.     

Microscopic characterization of petiole anatomy of Asteraceous taxa of Western Himalaya-Pakistan

Petiole anatomy of 15 species of family Asteraceae was examined which aimed to investigate petiolar anatomical structures for species level identification. Shandon Microtome was used for petiole histological preparations. Both qualitative and quantitative features were studied under microscope which showed significant variation in petiole, collenchyma, parenchyma shape/size, vascular bundles arrangement/size, and vessel elements quantity. Artemisia japonica Thunb., Cirsium vulgare (Savi) Ten., Myriactis nepalensis Less., Seriphidium brevifolium Ling & Y.R.Ling, Taraxacum officinale (L.) Weber ex F.H.Wigg., and Xanthium strumarium L. showed winged petioles. Maximum length and width of upper and lower epidermis was found in Tagetes erecta L. which is 23.05 ± 0.89 μm, 24.9 ± 1.257 μm length and 21.75 ± 1.38067 μm, 22.75 ± 0.467 μm width, respectively. Petioles of Parthenium hysterophorus L. was longest one with 9.85 ± 10.45 μm while A. japonica Thunb. showed highest number of vessel elements. Maximum size of vascular bundles was found in T. erecta L. with 5.05 ± 14.25 μm. Artemisia annua L., C. vulgare (Savi) Ten, Cyanthillium cinereum (L.) H.Rob., Helianthus annus L., M. nepalensis Less., P. hysterophorus L., Senecio chrysanthemoides DC. have trichomes while Tussilago farfara L. has highest number of vascular bundles. All species have angular collenchyma type except M. nepalensis Less., P. hysterophorus L., S. brevifolium Ling & Y.R.Ling, Tagetes minuta L., T. officinale L., S. chrysanthemoides DC., and T. farfara L. Cluster analysis implemented that distinct plant species in cluster. Petiolar anatomical structures and taxonomic key will helpful for distinguishing Asteraceous taxa at genus and species level. This taxonomic significant investigation will also provide baseline to taxonomists for other Asteraceae studies and phylogenetic research.     

Palyno‐morphological investigations of subtropical endangered flora of Capparidaceae through light and scanning electron microscopy

The research was performed to investigate pollen morphology of endangered species of Capparidaceae in subtropical regions of Pakistan. The distinguishing characters were investigated by using light microscope and scanning electron microscope. Palynological study is comprised of pollen shape, pollen type, exine sculpturing, polar and equatorial diameter, length and width of colpi, mesocolpium, and exine thickness. In polar view, Cleome viscosa exhibited the highest pollen size 26.4 (32.7–24.5 μm) ±0.776 whereas Capparis spinosa appeared to be the lowest 12.6 (14.5?10.7 μm) ±0.400. In equatorial view, Cleome viscosa had the largest pollen size 17.1 (20.0–15.0 μm) ±0.606 and Capparis spinosa had the smallest pollen size 9.7 (12.50–8.00 μm) ±0.394. The maximum fertility percentage has been observed in Capparis spinosa, that is, 98.96% and minimum in Cleome viscosa, that is, 82.93%. Diagnostic key has been constructed to state the essential diagnostic features by means of which the taxa can be identified. Remarkable variations have been observed in pollen size, shape, and exine sculpturing. All the selected species were tricolporate. Prolate to subprolate pollen were observed. There is a great variation existed in exine sculpturing such as in Capparis decidua and C. sp. nova sculpturing is reticulate, in Capparis himalayensis sculpturing is Scabrate granulate, in Capparis spinosa sculpturing is Psilate, in Cleome viscosa sculpturing is regulate‐reticulate, in Dipterygium glaucum sculpturing is regulate and in Gynandropsis gynandra sculpturing is striate‐regulate. On the basis of overall characteristics of pollen it seems that palynology of this family is helpful at the generic and specific level.     

Microscopic characters of the leaf and stem of Lavandula dentata L. (Lamiaceae)

Lavandula dentata L. is an aromatic plant used in folk medicine for different purposes and, for this reason, phytochemical surveys have been carried out in the search for bioactive substances aiming to support its uses. Since there is little knowledge on the structural aspects of L. dentata, this work has studied the anatomical characters of the leaf and stem using light and scanning electron microscopy, in order to assist the species identification. As a result, there are different types of trichomes: capitate glandular with uni‐ or bicellular head, peltate glandular with multicellular head, and branched non‐glandular. The leaf is hypostomatic showing diacytic stomata. The epidermis is uniseriate and coated with striate cuticle. The mesophyll is dorsiventral and the midrib is concave–convex and traversed by a single collateral vascular bundle. The stem is quadrangular and has alternating strands of collenchyma and cortical parenchyma as well as a typical endodermis in the cortex. The phloem and xylem cylinders are traversed by narrow rays and there is an incomplete sclerenchymatic sheath adjoining the phloem. These results are a novelty for the species and contribute to distinguish it from other lavenders. Microsc. Res. Tech. 77:647–652, 2014. © 2014 Wiley Periodicals, Inc.     

Comparative authentication of three “snow lotus” herbs by macroscopic and microscopic features

“Snow lotus” is a famous Chinese Materia Medica derived from species of the genus Saussurea (Compositae). To differentiate three representative easily‐confused snow lotus herbs, namely, Saussurea involucrata (Kar. et Kir.) Sch.‐Bip, Saussurea laniceps Hand.‐Mazz., and Saussurea medusa Maxim., macroscopic features of the three herbs were systemically observed, and microscopic features were compared by using ordinary light microscopy, polarized light microscopy and scanning electron microscopy (SEM). The results indicate that, as for macroscopic identification, capitula situation and arrangement, and as for microscopic identification, pollen grains, nonglandular hairs, glandular hairs, and cells of inner surface of the microdiodange can be used to authenticate the three snow lotus herbs. Comprehensive table comparing the characteristics were presented in this study. SEM has been found to provide a number of unique characteristics of pollen grains. Based on the observation of pollen grains, evolution sequence of the three species was speculated. The present method was proven to be efficient, convenient, simple, and reliable, which was successfully applied to the authentication of three snow lotus herbs. Microsc. Res. Tech.1 77:631–641, 2014. © 2014 Wiley Periodicals, Inc.     

Palyno-morphological attributes of some selected plant species of family Asteraceae from district Dera Ismail Khan,KPK, Pakistan

The current study aimed to investigate the palyno-morphological features of Asteraceous species from District Dera Ismail Khan, Khyber Pakhtunkhwa, Pakistan. A total of 13 species were collected, pressed, identified, and examined by using light and scanning electron microscopy. Both qualitative and quantitative pollen characters were observed, that is, equatorial and polar diameter, number of pores and colpi, exine thickness, exine sculpturing, pollen shape, and P/E ratio. Pollen shapes observed in studied taxa were spheroidal, suboblate, oblate-spheroidal, and lophate-spheroidal. The aperturation patterns of pollens vary from tricolporate to tetracolporate. The maximum polar and equatorial diameter of 75–100 (87.5 ± 13.6) μm and 87.5–117.5 (102.5 ± 16.4) μm, respectively, was recorded in Echinops echinatus, while minimum polar and equatorial diameter of 20–22.5 (21 ± 1.36) μm and 22.5–25 (23.5 ± 1.36) μm, respectively, was recorded in Aster subulatus. Six types of exine sculpturing patterns were observed; echinate, micro-echinate, echinate-microreticulate, microreticulate, echinate-perforate, and scabrate. Maximum exine thickness of 8.75 μm was recorded in E. echinatus and minimum of 1.25 μm in Launaea mucronata. The pollen morphology has valuable significance in understanding the taxonomy of different plant groups and plays a central role in the correct identification and classification of Asteraceous flora at species, generic, and tribe levels.     

Morphological,microscopic, and physicochemical studies of Diospyros montana

Adulteration is the root cause of producing not only a chemically and pharmacologically inferior but also in some instances hazardous or poisonous drug. Despite availability of several techniques, microscopy and physicochemical analyses are the most practical approaches for crude drug authentication. Hence, the present study aimed to evaluate morphological, microscopic, and physicochemical properties of root, bark, leaf, and fruit of Diospyros montana Morphological properties were determined by sensory organs, whereas microscopic features of cross‐sections and powders were determined by light and scanning electron microscopy. The proximate and fluorescence analyses were performed using the standard guidelines. The physical examination of fresh, shade‐dried, and powdered material showed no significant change in color. The identifying cellular structures included cuboidal cork, pitted tracheids, scalariform, reticulate and spiral xylary vessels, and rosettes, raphide, and cuboidal calcium oxalate crystals. The stomatal number, stomatal index, vein‐islet and vein‐termination number, and palisade ratio in the leaf were found to be 293.91 ± 32.68 mm^?2, 64.18 ± 3.42%, 22.00 ± 3.81 mm^?2 and 38.40 ± 5.81 mm^?2, and 3.85 ± 0.60, respectively. Total ash, acid insoluble ash, water soluble ash and sulfated ash of leaf (9.00 ± 0.50%, 1.67 ± 0.23%, 2.00 ± 0.22% and 14.50 ± 0.99%, respectively), foaming index of bark and root (111.11 ± 2.11), and swelling index of fruit (19.00 ± 3.45) were higher than the other parts. The powder of different parts showed characteristic colors in the daylight and UV light upon treatment with various regents. The plant was found to be rich in saponins, fibers, and flavonoids. The results of the present study may serve as identifiers of different parts of Diospyros montana.     

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.     

Microscopic and phytochemical techniques as a tool for authentication of herbal drug chiraita: Swertia cordata (G. Don) C.B. Clarke

Swertia cordata (G. Don) C.B. Clarke is one of the potential medicinal plants extensively used in eastern traditional medicine such as Unani, Ayurveda, Siddha, and in traditional Tibetan and Chinese medicine. S. paniculata is the common adulterant of S. cordata at herbal shops and markets but S. paniculata is also used in number of herbal formulations. The present study was conducted to use microscopic, pharmacognostic, and phytochemical techniques as a tool for the authentication of herbal drug chiraita (S. cordata). In herbal markets, mixing, adulteration, and use of spurious materials as substitute have become a major concern for herbal practitioners, local user, and industry for reasons of safety and efficacy. Therefore, authentication of medicinal plants is of utmost importance at each level of drug research. In the present study, anatomical features of two species showed a great diversity, as irregular epidermal cells and nonglandular, unicellular trichomes were found in S. cordata while in S. paniculata epidermal cells were hexagonal in shape and trichomes were A‐shaped. Antioxidant activity of two species showed a great variation where IC₅₀ value recorded for S. cordata was 208 μg/mL, while for S. paniculata IC₅₀ was 624 μg/mL. The study can serve as an important source of information to achieve the authenticity and to evaluate the quality and purity of the plant material in accordance to WHO guidelines. As this species is greatly exploited, so conservation is highly recommended.     

Microscopic investigations and pharmacognostic techniques used for the standardization of leaf of Rhus succedanea var. Himalaica J. D. Hook

Rhus succedanea is generally traded, distributed, and sold in the markets in its crude and raw form. This may have been mixed with adulterants, mismanaged by malpractices, and substituted with other closely related drugs having different effect. This study is therefore carried out to authenticate the plant through pharmacognostic evaluations including morphological microscopic and physio‐chemical features. Morphologically, the Rhus is a perennial small sized deciduous tree, locally, called as Rakhkal in Pashto and Kakarsingi in Urdu. The organoleptic evaluation showed that leaf had pleasant, aromatic odor, and astringent taste. Anatomy of the midrib has shown a typical dicot histological differentiation. The leaf was hypostomatic showing anomocytic stomata with average stomatal number 27.1 ± 7.2 and stomatal index 14 ± 3.63. The average vein islet, vein termination, and palisade ratios were 13.6 ± 3.04, 10.21 ± 1.92, and 6 ± 2.01, respectively. Leaf powder showed the existence of various types of tissues. Fluorescence study and percentage extractive values were also carried out. The phytochemical screening showed the presence of carbohydrates, protein, alkaloids, phenols, flavonoids, terpenoids, and anthraquinones, whereas tannins and fixed oil was not detected. Quantitatively highest number of alkaloids 16% and flavonoids 19% in leaf was detected. The above parameters, being reported to the first time for, and are significant toward establishing the microscopic and pharmacognostic standards for future identification and authentication of genuine herbal drug.