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.

Use of scanning electron microscopy to analyze sculpturing pattern and internal features of pollen grain wall in some members of Astragaleae (subfamily: Papilionoidae).

Palynological features of tribe Astagaleae L. have been observed with scanning electron microscopy (SEM), to assess features that can be used to re-examine the placement and taxonomic position of tribe. The palynomorph attributes of 10 species included in three genera of Astragaleae (Astragalus L., Glycyrrhiza L., and Oxytropis DC.) are investigated. The assignment of genus Oxytropis in tribe Astragaleae was also reevaluated based on pollen characters. Pollen appear to be prolate, prolate-spheroidal, and subprolate. Polar axis size (P) varies from 31.3 µm ±1 to 17.5 µm ±1.4 and equatorial axis size (E) varies from 22.2 µm ±1.8 to 13.1 µm ±0.9. Prominent apertures found consistently dispersed along the surface of pollen. Three kinds of ornamentation pattern were recorded, that is, reticulate or reticulate-perforate and perforate. The studied species display variation in equatorial and polar diameter, aperturation and sculpturing pattern, exine thickness, and pollen shape. On the basis of descriptive pollen features, a dichotomous taxonomic key and UPGMA analysis has been made for effortless and quick identification. The study concluded that disparities of the entire features are typically unrelenting and concerned with the species and shows potential systematic significance. The combination of palynological attributes in association with additional traits has prospective for systematic identification at species and genus level.

Application of spore morphology to solve identification problems in certain species of family Dryopteridaceae from Malakand Division, Pakistan.

Here we investigate the morphology of the spores of 12 native taxa of Dryopteridaceae that grow in Malakand division, Pakistan; most of these species not fully described before. This study intends to survey the taxonomic significance of spore morphological features and their variation useful for species identification in 7 species of Polystichum and 5 taxa of Dryopteris. The spores examinations were accomplished utilizing a light microscope (LM), and scanning electron microscope (SEM). The spores are monolete, medium-sized, with variable shapes in both equatorial and polar views. The mean estimation of equatorial diameter ranging from 28.3 to 58.3 µm and the polar diameter varied from 27.6 to 45.8 µm. The exospore thickness ranges between 1.6 to 3.8 µm, and the thickness of perispore is 0.8-6.7 µm. The perispore is perforated, ornamentation elements occur in different ways in the studied taxa of Polystichum. In investigated Dryopteris taxa, rugate perispore type is found. Results of multivariate statistical analysis (UPGMA, and PCA) established on quantitative and qualitative morphological traits of spores demonstrate that the species form distinct groups, not reflecting any phylogenetic relationships. A key to the spores types is provided for species identification, which dependent on the diagnostic characters of spores. The results of this study indicate that variation of spore morphological traits; in particular, the ornamentation and perispore characteristics, which is surface texture, have diagnostic value at the species level. Our data will help in the identification of spores of the family Dryopteridaceae in palynological and taxonomic studies.

Pollen diversity and its implications to the systematics of mimosaceous species by LM and SEM.

In this study we have discussed pollen morphology of 14 mimosaceous species belonging to five genera by using light microscopy (LM) and scanning electron microscopy (SEM). The palynological features of species were determined by both qualitative and quantitative characters. The qualitative characters include pollen shape, colpi arrangement, and exine sculpturing. Oblate spheroidal shaped pollens were noted in Albizia procera, Albizia lebbeck, Acacia tortilis, Acacia ampliceps, and Acacia modesta, subprolate shaped pollen in Leucaena leucocephala, Prosopis julifera, and Acacia nilotica, prolate pollen in Acacia farnesiana and Prosopis glandulosa while spheroidal, sub spheroidal and sub oblate pollen grains were observed in Acacia catechu, Mimosa himalayana, Prosopis cineraria, respectively. Sparsely foveolate, reticulate, scabrate, and scrobiclate exine sculpturing were observed. Colpi with sunken ornamentation were seen in P. cineraria, P. glandulosa, P. juliflora, and L. leucocephala while colpi were absent in rest of the studied species. The quantitative characters were statistically studied using SPSS software. The current study gives important morpho-palynological characters for identifying and validation of close related and similar taxa which will aid to the phylogenetic analysis of Mimosaceae family.

Seed morphology: An addition to the taxonomy of Astragaleae and Trifolieae (Leguminosae: Papilionoidae) from Pakistan.

Seed morphology was described in detail for 12 species belonging to 5 genera of tribes Astragaleae and Trifolieae (Leguminosae; Papilionoideae) using scanning electron microscopy (SEM) to evaluate the taxonomic relevance of macromorphological and micromorphological seed characters. The study aims to search for diagnostic seed ultrastructural features that may help to elucidate species identification. For SEM analysis, seed morphological characters including seed form and shape, color and size, ornamentation, epidermal cell shape, and anticlinal wall pattern were investigated. As a result of the study, species-specific characters have been determined. Based on seed exomorphology, three characteristic cell patterns; irregular, round, and flat were observed. In majority of studied taxa, species may be further differentiated based on seed shape, size, and surface ornamentation. The inconsistency in testa cell pattern, shape, and distribution of papillae or protuberances may probably give further insight and significant morphological features at specific and generic level within the tribe. This study illustrated that considerable taxonomic knowledge can be obtained by examining the seed characters of Astragaleae and Trifolieae, particularly at the species level. The results demonstrated that the use of SEM in seed morphology could play a role in the identification of taxa particularly at genus and species level. Twelve species of Astragaleae and Trifolieae were studied in order to describe and investigate the seed morphology and to evaluate the diagnostic value of this character using a SEM. A broader taxon sampling is required for classification at generic and tribal level, besides molecular and phylogenetic studies.

Spore morphology and leaf epidermal anatomy as a taxonomic source in the identification of Asplenium species from Malakand division Pakistan.

Several studies have demonstrated the usefulness of leaf epidermal, and spore morphological characters in the taxonomy of genus Asplenium. However, works on the Pakistani species of Asplenium are not existent. With the objective to verify the efficacy of leaf epidermal and spore morphological traits, the leaf epidermis and spore morphology in nine Asplenium species occurring in Malakand division was studied. The spores were studied under light microscope (LM) and scanning electron microscope (SEM), whereas for leaf epidermal anatomy, the LM was used. The spores are monolete, ellipsoidal in shape, the equatorial and polar diameter ranges between 28.3 and 50.2 × 27.6 and 45.8 µm. The exospore thickness varied from 0.5 to 3.8 µm. The perispore is 0.8-3.5 µm thick, ornamented, and morphologically variable. The spores characters specifically the perispore ornamentation are useful in distinguishing species within the genus. The irregular spore shape and unusual development of perispore wall surface characterize aborted spores in the species of Asplenium × alternifolium. The most informative quantitative characters appeared to be length and width of epidermal cells, and length and width of stomata were useful to distinguish species. The most significant qualitative characters to distinguish species were the anticlinal wall pattern. Our study has shown that considerable variations exist in the leaf epidermis of Asplenium species, at least some of which has taxonomic significance. We confirmed the prevalent taxonomic value of stomatal and epidermal cells traits. The leaf epidermal anatomy and spore morphological features showed to be a good source of information for taxonomy of the genus Asplenium.

An ethnobotanical study of medicinal plants used to treat skin diseases in northern Pakistan.

BACKGROUND: Skin diseases are a major health concern especially in association with human immune deficiency syndrome and acquired an immune deficiency. The aim of this study was to document the ethnomedicinal information of plants used to treat skin diseases in Northern Pakistan. This is the first quantitative ethnobotanical study of therapeutic herbs utilized by the indigenous people of Northern Pakistan for skin diseases. METHODS: Interviews were taken to obtain information from 180 participants. Quantitative methods including fidelity level (FL), Frequency of citation (FC), Use-value (UV), Jaccard indices (JI), Family importance value (FIV), Relative frequency of citation (RFC) and Chi-square test were applied. Medicinal plants uses are also compared with 50 national and international publications. RESULTS: In this study, we recorded 106 plant species belonged to 56 floral families for treatment of skin ailments. The dominant life form reported was herb while the preferred method of utilization was powder, along with leaf as the most used plant part. RFC ranges from 0.07 to 0.25% whereas the highest FIV was recorded for family Pteridaceae. FL values range from 36.8 to 100%. The study reported 88% of new plant reports for the treatment of skin diseases. CONCLUSION: The present study revealed the importance of several plants used to treat skin diseases by the local communities of Northern Pakistan. The available literature supported the evidence of plant dermatological properties. Plants having high UV and RFC can be considered for further scientific analysis. There is dire need to create awareness among local, government and scientific communities for the preservation of medicinal species and ethnomedicinal knowledge in Northern Pakistan.

Taxonomic importance of spore morphology in Thelypteridaceae from Northern Pakistan.

Spore morphology of Thelypteridaceae species growing in Malakand Division, Northern Pakistan, was studied using both light microscopy and scanning electron microscopy. The taxa are Christella dentata and Glaphyropteridopsis erubescens in the subfamily Thelypteridoideae, and Phegopteris connectilis, Pseudophegopteris pyrrhorhachis, and Pseudophegopteris levingei in the subfamily Phegopteridoideae. The studied species exhibit differences in spore size, exospore thickness, color, and ornamentation. Spores of the studied species are monolete and medium-sized, and shape is ellipsoidal in both polar and equatorial views. The average measurement of the polar diameter ranges from 27 µm to 31 µm, whereas in the equatorial direction it varied from 20 µm to 40 µm. The exospore thickness ranges from 1.2 µm to 2.4 µm. Reticulate, laevigate with microgranules, cristate, and coarsely echinate surface ornamentation are observed among the species. Multivariate analysis including unweighted pair group method with arithmetic mean and principal component analysis was used for the grouping and discrimination of species and genera.

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.

Ethnobotany of Anti-hypertensive Plants Used in Northern Pakistan.

Hypertension is one of the most important factors responsible for cardiovascular ailments worldwide. It has been observed that herbal products and alternative herbal therapies played a significant role in decreasing hypertension. The aim of the current study is to provide significant ethnopharmacological information, both qualitative and quantitative on medicinal plants related to hypertension from Northern Pakistan. The documented data were quantitatively analyzed for the first time in this area. A total of 250 participants were interviewed through semi-structured discussions and questionnaires. Quantitative indices including FC (Frequency citation), FIV (Family importance value), RFC (Relative frequency of citation) and DCI (Disease Consensus index) were calculated. A total of 192 plant species, belonging to 77 families were reported to be used in treatment of hypertension in Northern Pakistan. The most dominant life form reported was herbs (54%), with decoction (72 reports) and leaves (55.1%) were commonly utilized plant part. Highest FIV was recorded in Lamiaceae (327 FIV). RFC ranged from 0.08 to 1.08% while DCI varied from 0.233 to 0.000. In this study original data was compared with thirty one previous national and international published papers from neighboring region to compare the medicinal uses and obtain some novel plant species. About 42% of the medicinal plant species were reported for the first time in treatment of hypertension in comparison to these 31 published papers. Different phytochemical activities of antihypertensive plants were also reported from literature. This research work documents the traditional knowledge of medicinal plants usage and provides baseline in designing clinical trials and pharmacological analysis for treatment of hypertension.

Foliar epidermal micromorphology and its taxonomic implications in some selected species of Athyriaceae.

For the robust identification of taxonomically complex fern family like Athyriaceae, light and scanning electron microscopy is significance implications. This article present first microscopic investigation of foliar micromorphology of 3 genera and 10 species belonging to Athyriaceae namely, Athyrium, Deparia, and Diplazium were collected from different localities in Malakand Division, Northern Pakistan. In present study we compare foliar micromorphology of all 10 species using standard protocols of light microcopy (LM) and scanning electron microscopy. Qualitative micromorphological variations in shape of epidermal cells, anticlinal wall pattern, stomatal type and shape, stomatal pore shape, guard cells shape, and trichomes types were studied. In addition, some quantitative characters were also studied and data were statistically analyzed in epidermal cell size, stomatal size, stomatal pore size, stomatal density, and stomatal index. The pivotal result of study include; shape of epidermal cell in all species is irregular on both abaxial and adaxial surfaces. The anticlinal walls are sinuous in most of the species but some species have irregular lobed and broadly lobed wall. Leaves are hypostomatic in all studied species. Two main categories of stomatal type were found: polocytic and anomocytic. Unicellular nonglandular trichomes were observed in only one species Athyrium mackinnoni. The variation in foliar micromorphological characters between the genera and within the species was useful in identification and classification and have potential taxonomic significance for species differentiation. An identification key using micromorphological characters are provided to distinguish genera and species.

A light and scanning electron microscopic diagnosis of leaf epidermal morphology and its systematic implications in Dryopteridaceae: Investigating 12 Pakistani taxa.

Dryopteris and Polystichum are the 2 complex taxonomic genera of Dryopteridaceae. The comparative foliar epidermal anatomy of 12 species of both genera from Pakistan were studied using standard protocols of light microscopy (LM) and scanning electron microscopy (SEM). The objective of which was systematic comparison and investigation to elucidate the taxonomic importance of foliar micromorphology, which may be useful to taxonomists for identifying complex Dryopteridaceae taxa. Principal component analysis and UPGMA clustering analysis were performed to test the validity of leaf anatomical features as method of separating species and genera. The foliar epidermal anatomy described here is a good source of taxonomic characters in both groups that can help genera and species delimitation. This is the first report on leaf micromorphology in most of these species. Observation of foliar anatomy showed that stomata are only present on the abaxial surface; i.e., leaves of all species are hypostomatic. The shapes of epidermal cells in all studied species are irregular. The anticlinal walls are strongly lobed, irregular wavy and elongated wavy. However, substantial variation in epidermal cell size and other stomatal features were observed on both upper and lower surfaces in all investigated species. Two types of stomata were observed in all studied species. The presence of polocytic stomata in Dryopteris and staurocytic stomata in Polystichum are the important characters for the segregation of these genera. Elongate elliptic stomatal shape, narrow kidney shaped guard cells and broad elliptic shaped stomatal pores are diagnostic for all five species of Dryopteris selected. On the other hand, size and number of epidermal cells, lobes per cell, stomatal size, subsidiary cell size, stomatal pore size and stomatal index are the key features for species differentiation in Polystichum. An identification key was developed in order to apply the foliar anatomical characters in the discrimination of the species studied.

Botany, ethnomedicines, phytochemistry and pharmacology of Himalayan paeony (Paeonia emodi Royle.).

ETHNOPHARMACOLOGICAL RELEVANCE: Himalayan paeony (Paeonia emodi Royle.) is an important species used to treat various diseases. This study aimed to compile the detailed traditional medicinal uses, phytochemistry, pharmacology and toxicological investigations on P. emodi. This study also highlights taxonomic validity, quality of experimental designs and shortcomings in previously reported information on Himalayan paeony. METHODS: The data was extracted from unpublished theses (Pakistan, China, India and Nepal), and different published research articles confined to pharmacology, phytochemistry and antimicrobial activities using different databases through specific keywords. The relevant information regarding medicinal uses, taxonomic/common names, part used, collection and identification source, authentication, voucher specimen number, plant extracts and their characterization, isolation and identification of phytochemicals, methods of study in silico, in vivo or in vitro, model organism used, dose and duration, minimal active concentration, zone of inhibition (antimicrobial study), bioactive compound(s), mechanism of action on single or multiple targets, and toxicological information. RESULTS: P. emodi is reported for diverse medicinal uses with pharmacological properties like antioxidant, nephroprotective, lipoxygenase inhibitory, cognition and oxidative stress release, cytotoxic, anti-inflammatory, antiepileptic, anticonvulsant, haemaglutination, alpha-chymotrypsin inhibitory, hepatoprotective, hepatic chromes and pharmacokinetics of carbamazepine expression, ß-glucuronidase inhibitory, spasmolytic and spasmogenic, and airway relaxant. Data confined to its taxonomic validity, shows 10% studies with correct taxonomic name while 90% studies with incorrect taxonomic, pharmacopeial and common names. The literature reviewed, shows lack of collection source (11 reports), without proper source of identification (15 reports), 33 studies without voucher specimen number, 26 reports lack information on authentic herbarium submission and most of the studies (90%) without validation of taxonomic names using recognized databases. In reported methods, 67% studies without characterization of extracts, 25% lack proper dose, 40% without duration and 31% reports lack information on proper controls. Similarly, only 18% studies reports active compound(s) responsible for pharmacological activities, 14% studies show minimal active concentration, only 2.5% studies report mechanism of action on target while none of the reports mentioned in silico approach. CONCLUSION: P. emodi is endemic to Himalayan region (Pakistan, China, India and Nepal) with diverse traditional therapeutic uses. Majority of reviewed studies showed confusion in its taxonomic validity, incomplete methodologies and ambiguous findings. Keeping in view the immense uses of P. emodi in various traditional medicinal systems, holistic pharmacological approaches in combination with reverse pharmacology, system biology, and "omics" technologies are recommended to improve the quality of research which leads to natural drug discovery development at global perspectives.

Traditional knowledge on herbal drinks among indigenous communities in Azad Jammu and Kashmir, Pakistan.

BACKGROUND: Traditional knowledge about the use of medicinal plants for herbal drinks (HDs) is not well documented in the Azad Kashmir region despite their widespread use. This study highlights the taxonomic diversity and traditional knowledge on medicinal plants used for HDs while examining the diversity of diseases treated with HDs in the study area. METHODS: Individual discussions were conducted with 255 informants (84 women and 171 men). Data gathered included (i) informant age and gender, (ii) HD species and respective plant parts used, (iii) health disorders treated, and (iv) mode of preparation and utilizations. Quantitative ethnobotanical indices including relative frequency of citation (RFC), informant consensus factor (ICF), and use value (UV) were used for data analyses. RESULTS: Altogether, 73 medicinal plants belonging to 40 families and 66 genera were reported to be used in HD preparations, with Asteraceae being the richest family. The average number of HD species cited was 9.09 ± 0.17 per informant and did not vary either by age or gender. In addition, men and women, and adults and the young used the same pool of species (dissimilarity nearly zero). The most used plant parts were leaves (20.00%), roots (17.25%), and fruits (16.47%). Based on UV, the top five most used species were Valeriana jatamansi, Isodon rugosus, Onopordum acanthium, Acacia nilotica, and Viola canescens; and the UV was similar among gender and age categories too. The most utilized herbal preparation forms included decoctions, infusions, and tea. One hundred and eleven diseases grouped into 13 ailment categories were reported to be cured using HDs. The main category of disease treated with HDs was gastrointestinal (GIT) disorders (RFC = 17.43%). Relatively few species were used by a large proportion of informants for each category of ailment (ICF ≥ 0.60). Only one species was used for "glandular disorders" and "eye diseases" (ICF = 1). A novelty of about 22% (16 species) was recorded for HD species in the present work. CONCLUSION: The diversity of medicinal plant species used as HDs and the associated traditional knowledge are of considerable value to the indigenous communities of the Azad Kashmir region. Therefore, there is a need for conservation and preservation of medicinal HD species as well as the wealth of indigenous knowledge. The conservation effort should be high for species in the ailments categories glandular disorders and eye diseases. The therapeutic uses of HDs have provided basic data for further research focused on phytochemical and pharmacological studies and conservation of the most important species.

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

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