Morpho-palynological investigation of gymnospermal flora from subalpine and alpine zones of northern Pakistan using LM and SEM.

The pollen morphology, with special reference to exine sculpture, of some species of the gymnosperms was assessed for the first time from the subalpine and alpine zones of western Himalayas northern Pakistan. The pollen of all these species is airborne and allergenic, so pollen morphology helps for identification of this allergenic pollen at specific level. Different morpho-palynological characteristics were analyzed including size range of pollen, polar and equatorial diameter ratio, exine ornamentation, sculpturing, exine thickness, pollen type, and shape. For accurate and quick identification of species, taxonomic key was made based on different morpho-palynological characteristics. The quantitative data were processed using SPSS software. Gymnospermal pollen includes inaperturate, rarely 1-colpate observed in (Cupressaceae), hexazonocolpate in (Ephedraceae), vesiculate, bissacate in (Pinaceae), and inaperturate in (Taxaceae). Different pollen shapes observed were prolate (4 spp), sub-spheroidal (7 spp), and oblate (1 spp). Variation was observed in exine sculpturing granular (4 spp), reticulate (1 spp), areolate-punctate (3 spp), and psilate (2 spp). This is based on the analysis of 10 plants belonging to four families of gymnosperms. Distinct pollen shape has emerged as the most diagnostic feature to separate some genera such as spheroidal in (Cupressaceae, Taxaceae), prolate and radiosymmetrical in (Ephedraceae), and bilateral in (Pinaceae). Exine thickness and sculpturing proved to be helpful at generic and specific levels. The results reinforced the significance of gymnospermal pollen morphological features which were used as aid for valuable taxonomic tool in plant systematics.

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.

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.

Comparative light and scanning electron microscopy in authentication of adulterated traded medicinal plants.

The medicinal plants are utilized globally considering the cheap and chemical free source, but their correct identification and authentication is prerequisite for safety and efficacy of plant-based medicines. The present study encompassed traded medicinal plants (16) with high therapeutic value from diverse families like Brassicaceae, Berberidaceae, Malvaceae, Salicaceae, Myrtaceae, Papilionaceae, Ascelpiadaceae, Colchicaceae, Violaceae, and Vitaceae for detailed microscopic study of characters that is, morphology, pollen shape and sizes, P/E ratio, pore length and width, spine length, colpi dimensions, and exine sculpture pattern. The plants showed noteworthy differences in microscopy of Wattakaka volubilis having pollinia, translator and corpusculum like structures while pores were visible in Colchicum luteum, Alcea rosea, and Hibiscus syriacus. The spines were observed in Centipeda minima, A. rosea, and H. syriacus being dimorphic spines in A. rosea and monomorphic in H. syriacus. The exine sculpturing pattern was reticulate in mostly studied plants however distinctive exine pattern was noted in Berberis aristata and Berberis lyceum. The highest polar diameter, equatorial diameter and exine thickness among studied plants were observed in H. syriacus (161 µm), C. luteum (50 µm) and Vitis jacquemontii (1.10), respectively. Thus, microscopy of medicinal plants in addition to other taxonomic evidence offers a supportive skill in authentication, consequently utilization by local consumers and pharmaceutical industries.

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.

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.

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.

Microscopic investigations and pharmacognostic techniques used for the standardization of herbal drug Nigella sativa L.

A comprehensive account on standardization of herbal drug Nigella sativa L. (Tukhm-e-Kalonji) by using microscopic as well as pharmacognostic parameters. In the field of herbal medicines, the main issues are quality, purity, and effectiveness, as in many cases herbal drugs are knowingly or unknowingly substituted or adulterated with similar species or varieties. In herbal market the seeds of N. sativa are commonly adultered with seeds of Allium cepa L. intentionally or accidentally due to their similar morphology. In the present study, the microscopic characterization of herbal drug was done by morphological, palynological, and anatomical features. A great diversity was found in anatomical features of two species, as irregular epidermal cells, actinocytic stomata, and nonglandular trichomes were found in N. sativa while in A. cepa epidermal cells were rectangular in shape, stomata type was paracytic, and trichomes were absent. Pharmacognostic characterization was made by fluorescence analysis and physiochemical parameters. Physicochemical parameters like moisture content, total ash, acid insoluble ash, water soluble ash, and water insoluble ash were also evaluated. The above parameters, being reported to the first time for the studied plant species, and are significant towards establishing the microscopic and pharmacognostic standards for future identification and authentication of genuine herbal drug. RESEARCH HIGHLIGHTS: Microscopic Investigations for the standardization of herbal drug Nigella sativa L. (Tukhm-e-Kalonji). Pharmacognostic techniques are potentially significant for the standardization of herbal drug Nigella sativa in comparison with its adulterant. Useful for preparation of standards for herbal pharmacopeia.

Ethnopharmacological evaluation and antioxidant activity of some important herbs used in traditional medicines.

OBJECTIVE: To document the indigenous knowledge and evaluate the antioxidant activity of medicinal herbs used for treatment of various diseases. METHODS: The ethnomedicinal data were collected through semi-structured interviews of local informants along with collection of herbarium specimens. The antioxidant activity was evaluated by using 1-diphenyl-2-picryl-hydrazyl radical scavenging assay. RESULTS: A total of 12 important medicinal herbs were evaluated for ethnomedicinal uses and antioxidant activity. Medicinal plant diversity showed that Solanaceae was the dominating family (3 spp.), followed by Asteraceae and Lamiaceae (both 2 spp.). Leaves (31% ) were most frequently used in preparation of traditional medicines, and the most recommended mode of preparation was powder (28% ). The antioxidant results revealed that the highest activity was found in Atropa acuminata and Crocus sativus. CONCLUSION: The results of this study proved that A. accuminata and C. sativus can be the excellent sources of antioxidant compounds. A sustainable use and conservation of the valuable natural resource greatly promote the development of functional food additives and cosmetics.

Ethnobotanical survey of medicinally important shrubs and trees of Himalayan region of Azad Jammu and Kashmir, Pakistan.

ETHNOPHARMACOLOGICAL RELEVANCE: Present study was commenced with an aim to document the indigenous knowledge of medicinally important shrubs and trees of Himalayan region of Azad Jammu and Kashmir, Pakistan. This is the first contribution to the quantitative ethnobotany of this region, as no reported data focusing on shrubs and trees from the area have been published. Study reported the ethnobotanical significance of medicinal plants for the treatment of various diseases. METHOD: Study was conducted during 2012-2014 following standard ethnobotanical methods. The ethnomedicinal data was collected through informed consent semi- structured interviews of 160 key informants. Documented data was analyzed by using quantitative indices of informant consensus factor (ICF), fidelity level (FL), use value (UV) and relative frequency citation (RFC). RESULTS: A total of 73 shrub and tree species belonging to 56 genera and 37 families were reported to be used ethnomedicinally for the treatment of various ailments. Medicinal plant diversity showed that Rosaceae was dominating family with (9 spp.) followed by Moraceae (7 spp.), Euphorbiaceae, Mimosaceae, Pinaceae, Rhamnaceae (4 spp. each), Oleaceae (3 spp.), Apocynaceae, Caesalpinaceae, Ebenaceae, Fagaceae, Lythraceae, Papilionaceae, Acanthaceae, Verbenaceae (2 spp. each) while remaining 22 families were represented by one species each. Leaves (23%) were highly utilized plant parts, followed by fruits (22%), bark (18%), seeds (10%), roots (9%), flowers (8%), whole plant and aerial parts (4% each) and stem (2%). Modes of preparation fall into 14 categories including powder (33 reports) followed by decoction (29 reports), paste (22 reports), juice (18 reports), infusion (12 reports), raw (8 reports), extract and latex (5 reports each), gum and oil (4 reports each), fresh part and pulp (2 reports each), chewed and cooked (1 report each). The highest FIC was recorded for Gastro-intestinal disorders (0.58) followed by nail, skin and hair disorders (0.44). Maximum fidelity level (FL=100%) was expressed by Abies pindrow, Adhatoda vasica, Bauhinia variegata and Cedrela serrata. Based on use value Juglans regia (0.88) was found most significant species followed by Acacia nilotica (0.83), Phyllanthus emblica (0.81), Pinus roxburghii (0.75) and Punica granatum (0.71). CONCLUSION: The area has a rich diversity of medicinally important shrub and tree species. The tradition of using plants for medicinal purposes is still alive in the local community but recently this tradition is gradually declining in new generation. Therefore awareness is needed to be raised among the local people on sustainable use and conservation of local flora.

Ethnomedicinal knowledge of the most commonly used plants from Deosai Plateau, Western Himalayas, Gilgit Baltistan, Pakistan.

ETHNOPHARMACOLOGICAL RELEVANCE: The current survey was aimed to identify the prime important medicinal plants in terms of medicines and further pharmacological screening of such plants. As far as, we know, no reported data from Deosai Plateau have been published and this is the first documented information of the study area. MATERIAL AND METHODS: Ethnomedicinal information of plants was collected through semi-structured interviews through a participatory rural appraisal (PRA) method. RESULTS AND DISCUSSION: This study provided information of 50 medicinal plant species used to treat 29 different ailments. A total of 42 species (84%) were herbs while shrubs and subshrubs were represented by 8 species (16%). Leaves (37%) were the predominant plant parts followed by, roots (27%), aerial parts (13%), flowers (12%), fruit (7%), seeds (3%), and tuber (1%). Method of preparation fall into five categories including infusion (31%), paste (23%), decoction (20%), powder (18%), and juice (8%). CONCLUSION: The study area is rich in medicinal plant diversity growing in wild. Our efforts in this regard are only a little contribution to the ethnobotanical study of this area focusing on medicinal plants while more clinical studies are required in future to prove such claims of local inhabitants.

An ethnobotanical study of medicinal plants in high mountainous region of Chail valley (District Swat- Pakistan).

BACKGROUND: This paper represents the first ethnobotanical study in Chail valley of district Swat-Pakistan and provides significant information on medicinal plants use among the tribal people of the area. The aim of this study was to document the medicinal uses of local plants and to develop an ethnobotanical inventory of the species diversity. METHODS: In present study, semi-structured interviews with 142 inhabitants (age range between 31-75 years) were conducted. Ethnobotanical data was analyzed using relative frequency of citation (RFC) to determine the well-known and most useful species in the area. RESULTS: Current research work reports total of 50 plant species belonging to 48 genera of 35 families from Chail valley. Origanum vulgare, Geranium wallichianum and Skimmia laureola have the highest values of relative frequency of citation (RFC) and are widely known by the inhabitants of the valley. The majority of the documented plants were herbs (58%) followed by shrubs (28%), trees (12%) and then climbers (2%). The part of the plant most frequently used was the leaves (33%) followed by roots (17%), fruits (14%), whole plant (12%), rhizomes (9%), stems (6%), barks (5%) and seeds (4%). Decoction was the most common preparation method use in herbal recipes. The most frequently treated diseases in the valley were urinary disorders, skin infections, digestive disorders, asthma, jaundice, angina, chronic dysentery and diarrhea. CONCLUSION: This study contributes an ethnobotanical inventory of medicinal plants with their frequency of citations together with the part used, disease treated and methods of application among the tribal communities of Chail valley. The present survey has documented from this valley considerable indigenous knowledge about the local medicinal plants for treating number of common diseases that is ready to be further investigated for biological, pharmacological and toxicological screening. This study also provides some socio-economic aspects which are associated to the local tribal communities.