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.     

Foliar epidermal anatomy of some selected wild edible fruits of Pakistan using light microscopy and scanning electron microscopy

The present study is focused on the detailed foliar epidermal anatomy of some selected wild edible fruits (WEFs) from Pakistan using light microscopy (LM) and scanning electron microscopy (SEM). The studied species are Ficus racemosa L., Solanum nigrum L., Capparis spinosa L., Physalis divaricata D.Don, Rosa moschata Herrm. and Ribes orientale Desf. collected from various localities of Pakistan. The objective of the present study is to investigate qualitative and quantitative anatomical characters for the identification and differentiation of collected wild edible fruits. The characters studied are shape and size of epidermal cells, anticlinal wall pattern, trichome type and shape, average number of stomata, length and width of stomata and pore. The detailed microscopic investigation and variations in the characters recorded have a key role in the determination and authentication of wild edible fruits. This study possesses great potential for plant taxonomists to further evaluate the species at molecular and genetic levels.     

Quality assurance and authentication of herbal drug (Argyrolobium roseum and Viola stocksii) through comparative light and scanning electron microscopy

The quality assurance and authentication of crude herbal drugs play important role in the effective therapeutic effect of herbal drug and their products. There are many reported problems in quality assurance of herbal crude drugs concerning to their correct identification. The present study was designed with the aim to document the authentication and quality assurance of the herbal crude drugs (Argyrolobium roseum and Viola stocksii) thorough light microscopy (LM) and scanning electron microscopy (SEM). The detailed foliar anatomical studies showed polygonal epidermal cells having anticlinal walls in Argyrolobium roseum while rounded epidermal cells were observed in Viola stocksii. The anomocytic stomata type was observed in Argrolobium roseum while actinocytic was noticed in Viola stocksii. The pollen of studied species appeared as tricolporate showing reticulate exine sculpturing in Argrolobium roseum while fine perforations were recorded in Viola stocksii. Furthermore, quantitative analysis of phenols, flavonoids, and antioxidant activity showed high flavonoid and phenol content in Argyrolobium roseum as compared with Viola stocksii. It was observed that Argyrolobium roseum was discriminated from the Viola stocksii based on the leaf and pollen micromorphological traits by using LM and SEM techniques. It was concluded that LM and SEM techniques were found useful for the quality assurance of botanicals and their authentication.     

Light microscopy and scanning electron microscopy: Implications for authentication of misidentified herbal drugs

Plant‐based drugs have reached remarkable acceptability as therapeutic remedy for various diseases due to the adverse effects of contemporary medicines. This increasing popularity of herbal drugs leads to a growing herbal market for the development of plant‐based drugs, nutraceuticals, pharmaceuticals and cosmeceuticals. Herbal drug adulteration is a complex problem which currently has undeniable consequences on health and nutrition. Ambiguities in nomenclature, misidentification and resemblance of colour and texture of the crude herbal drugs are the major causes of adulteration. Three different species commercially marketed under the same trade name Halion are Lepidium apetalum, Asparagus officinalis, and Lepidium didymum. The genuine source of Halion is Lepidium apetalum, which is authenticated by using basic and advanced taxonomic techniques. Morphology, anatomy and palynology of the misidentified sources were done using light and scanning electron microscopic techniques for authentication. This study may help to set microscopic techniques as a tool to achieve quality and standardization of the genuine source of the herbal drug. Phytochemical analysis and biological screening is needed for the further establishment of authenticity and quality of herbal drugs.     

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.     

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.     

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.     

Detailed macro and micromorphology of grasses caryopsis using scanning electron and light microscopy

Micromorphology of 15 grass caryopsis was investigated through scanning electron microscopy for its implications in the identification of grasses. Both macro and micromorphological characters especially caryopsis shape and surface morphology were found significant in the taxonomic diagnosis. Shape morphology of caryopsis varied among the species and lanceolate, fusiform, narrow ellipsoidal, ovate, elliptical, plano-convex, fusiform, linear ovate, broadly elliptical, broadly pllipsoidal, plano-convex, oblique ovate, and subglobose shapes were recorded. Digitaria nodosa (3.31 × 0.7 mm) and Echinochloa crus-galli (3.26 × 1.44 mm) had the largest caryopsis size, while Acrachne racemosa (0.76 × 0.54 mm) and Eragrostis minor (0.62 × 0.38 mm) had the smallest caryopsis size. Macro and micromorphological characters were useful for the diagnosis of genus and species. Utilizing both qualitative and quantitative character taxonomic key at generic and species level were made.     

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.     

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.     

Authenticating and distinguishing the eight species of traditional Tibetan medicine “Meiduoluomi” by microscopic technique

Because of the morphological and macroscopic similarity, many species of Erigeron and Aster (Asteraceae) are confusable and usually used under the same name “Meiduoluomi” in traditional Tibetan medicine (TTM). To find an easy, quick, and reliable method to authenticate and distinguish the eight main medicinal plants of these species, the light microscope was used to reveal the morphoanatomic details. The fixed, sectioned, and stained plant materials and epidermis materials were studied by microscopic techniques. The results of the microscopic features are systematically described and illustrated, and comparison parameters are presented. Furthermore, a key to the eight species of “Meiduoluomi” was constructed. Microscopy can be unambiguously used to authenticate and distinguish the eight main species of TTM “Meiduoluomi.” Microsc. Res. Tech., 2009. © 2009 Wiley‐Liss, Inc.     

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.     

Authentication of herbal drug Tukhm‐e‐balango (Lallemantia royleana Benth.) using microscopic,pharmacognostic, and phytochemical characterization

The study is aimed to provide a comprehensive account on authentication of herbal drug named as Tukhm‐e‐balango (Lallemantia royleana Benth.) from the seeds of Ocimum basilicum by using microscopic, pharmacognostic, and phytochemical characterization. The crude medicinal plants and their parts are often adulterated or substituted in market due to improper identification by the consumers while among herbal plant sellers, taxonomic confusion is caused due to morphological similarities of the plant parts and lack of a standard identification system.In microscopy, both herbarium and fresh specimens were studied using qualitative and quantitative morphological characteristics of leaves, seeds, and pollen. For pharmacognosy, solubility, fluorescence, and physicochemical characterizers were analyzed whereas a total phenolic and flavonoids contents was determined in addition to DPPH radical scavenging activity. In current study, microscopic, pharmacognostic, and phytochemical characterization clearly differentiated L. royleana from O. basilicum. The major problem in herbal drug industry is caused due to confusion and controversy of certain synonyms used for more than one or two drugs. Sometimes, under the same common or local name, entirely different taxa are being sold in herbal markets. It is concluded that correct and proper identification of medicinal plants is very crucial to ensure the safety and efficacy of herbal medicines, as many medicinal plants are intentionally or unintentionally adulterated with similar species or varieties. In herbal market, the seeds of L. royleana are adulterated with seeds of O. basilicum due to their similar morphology.     

Significance of leaf morphoanatomical markers for the authentication of adultered drugs sold in herbal markets of district Lahore,Pakistan

A number of herbal plants sold in herbal markets of Lahore are under adulteration threat which can pose harmful health effects to end‐user. This adulteration problem of medicinal plants can be resolved by the implication of some valuable taxonomic parameter such as leaf epidermal anatomical characteristics. Hence, this research was aimed to provide viable anatomical markers in order to resolve this adulteration issue persisted in some common marketed medicinal plants of district Lahore, that is, Cinnamomum verum Presl., Cinnamomum tamala (Buuch.‐Ham.) T.Nees&Eberm., Gymnema sylvestre (Retz.) R.Br.ex Sm., Sphaeranthus indicus Linn., Artemisia maritima Linn., Achillea millifolim L., Adhatoda vasica Nees, Butea monosperma (Lam.) Taub, and Morus nigra L. Overall multiple anatomical variations (epidermal cell shape, their length and width, type of stomata, length and width of guard cells along with presence or absence of trichomes) had been reported in the study that could be worthwhile for the correct identification of medicinal plants. Irregular shapes of epidermal cells were observed in Cinnamomum verum and Achillea millifolium while pentagonal and polygonal cells were found in their adultaerants, that is, Canella winterana and Adhatoda vasica, respectively. Types of stomata were also strikingly varied among genuine plant and its adulterant, for example, anisocytic stomata were observed in Artemisia maritima while in its adulterant (Artemisia absinthium) anomocytic stomata were found. Similarly, paracytic stomata were observed in Butea monosperma, whereas its adulterant plant (Averrhoa carambola) characteristically possessed anisocytic stomata. Hence, anatomical characteristics were proved to be a valuable taxonomic tool in resolving the adulteration issue of medicinal plants.     

Microscopic investigations and pharmacognostic techniques for the standardization of Caralluma edulis (Edgew.) Benth. ex Hook.f.

Herbal medicines frequently suffer with quality controversies because of similar species or varieties. This often leads to sophistication or admixture of the crude drug as they share various look alike physical features. Commercially, stalks of Caralluma edulis are commonly adulterated with other species because of their analogous morphology. In the present research, the microscopic standardization of the drug was performed by structural, histological, and anatomical features. Pharmacognostical characterization was carried out by physicochemical (moisture content, swelling index, pH value, ash values, and extractive values) and fluorescence analysis. The chemical characterization by UV–Visible, Fourier transform infrared spectroscopy, and high‐performance liquid chromatography was also performed that disclosed the chief phytoconstituents of the herbal drug. The above mentioned constraints, being reported for the first time in C. edulis, are substantial in establishing the microscopic and pharmacopoeial standards for future identification and authentication.     

Foliar epidermal anatomical characteristics of taxa of Iris subg. Scorpiris Spach (Iridaceae) from Turkey

Iris L. is one of the important genus of family Iridaceae, consist of 56 taxa naturally occurred in Turkey. The similarities and variations in the subgenus overlapping the taxonomic positions of the species in the subgenera and needs anatomical assessment especially by microscopic techniques. In this study, the taxonomic significance of leaf anatomical characters of 10 Iris subgenus Scorpiris taxa were studied in detail and the relationship among these taxa were evaluated using microscopy techniques. Fresh leaf samples of species were fixed in 70% alcohol solution for anatomical observation under microscope. Eleven different micromorphological features were statistically analyzed to delimit the species in subgenus. Based on morphological and anatomical similarities, we studied relationships among; (1) ssp. turcica, ssp. caucasica, I. nezahatiae and I. pseudocaucasica; (2) correlation between ssp. turcica and ssp. caucasica; (3) association of I. galatica, I. persica, ssp. margaretiae and ssp. stenophylla with each other; (4) relationship between ssp. stenophylla and ssp. margaretiae; and (5) relevance between I. aucheri and I. peshmeniana. Moreover, the taxonomy of subgenus Scorpiris has been discussed in detail with novel and diagnostic features based on micromorphological physiognomies. We found that four species in this study are endemic to Turkey, while seven are critically endangered geophytes in the country. The leaf anatomical characteristics of 10 taxa were divided into three groups. Main aim of this research was to study the taxonomy of the complex subgenus Scorpiris through microscopic techniques.     

Systematic significance of pollen morphology and foliar epidermal anatomy of medicinal plants using SEM and LM techniques

Alternative medicinal source is a hot issue within medicine community now a days due to various side effects of allotropic drugs; most importantly the multidrug resistance phenomena. Medicinal plants provide an attractive and effective alternative to modern synthetic drugs as they contain different types of bioactive compounds, having great potential in treatment of various diseases. The present study deals with identification of micromorphological characteristics of 10 ethnomedicinally important plants of Northern Pakistan, that is, Aster himalaicus C. B. Clarke, Artemisia maritima (Berg.) Wellkomm, Isodon rugosus (Wall. ex Benth.), Myricaria squamosal Desv, Pedicularis groenlandica Retz. Polygonum aviculare L, Salvia himmelbauer E. Peter, Swertia kingie Hook. f., Tanacetum dolichophyllum (Kitam) Kitam, Verbascum epixanthinum Boiss. & Heldr. as potential source for pharmaceutical industry. The characters studied were shape and size of pollen, polar diameter and equatorial diameter, epidermal cell shape and size, stomata width and length, trichome shapes, and type. Most of the species had tricolporate pollen type but hexacolporate pollen was also observed in Isodon rugosus, Polygonum aviculare, Salvia himmelbauer. Different types of exine sculpturing observed were foveolate, scabrate, lophate, regular, steriate, and alveolate. Minimum equatorial diameter was noted in Tanacetum dolichophyllum (12.4 μm) and maximum in Myricaria squamosal (20.9 μm). Shape of epidermis cell ranged from rectangular and irregular, polygonal and pentagonal. Different types of stomata like anisocytic, actinocytic, paracytic, and diacytic were noticed. Variations in microscopic characters observed in the current study have great potential to be used as identification tools by the taxonomists for further studies.     

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.     

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.