Folk Knowledge and Perceptions about the Use of Wild Fruits and Vegetables-Cross-Cultural Knowledge in the Pipli Pahar Reserved Forest of Okara, Pakistan.

Wild fruits and vegetables (WFVs) have been vital to local communities for centuries and make an important contribution to daily life and income. However, traditional knowledge of the use of wild fruits is at risk of being lost due to inadequate documentation. This study aimed to secure this knowledge through intermittent field visits and a semi-structured questionnaire. Using various ethnobotanical data analysis tools and SPSS (IBM 25), this study identified 65 WFV species (52 genera and 29 families). These species, mostly consumed as vegetables (49%) or fruits (43%), were predominantly herbaceous (48%) in wild and semi-wild habitats (67%). 20 WFVs were known to local communities (highest RFC), Phoenix sylvestris stood out as the most utilized species (highest UV). Surprisingly, only 23% of the WFVs were sold at markets. The survey identified 21 unique WFVs that are rarely documented for human consumption in Pakistan (e.g., Ehretia obtusifolia, Euploca strigosa, Brassica juncea, Cleome brachycarpa, Gymnosporia royleana, Cucumis maderaspatanus, Croton bonplandianus, Euphorbia prostrata, Vachellia nilotica, Pongamia pinnata, Grewia asiatica, Malvastrum coromandelianum, Morus serrata, Argemone mexicana, Bambusa vulgaris, Echinochloa colonum, Solanum virginianum, Physalis angulata, Withania somnifera, Zygophyllum creticum, and Peganum harmala), as well as 14 novel uses and five novel edible parts. Despite their ecological importance, the use of WFVs has declined because local people are unaware of their cultural and economic value. Preservation of traditional knowledge through education on conservation and utilization could boost economies and livelihoods in this and similar areas worldwide.

Cross-cultural ethnobotany of the Baltis and Shinas in the Kharmang district, Trans-Himalaya India-Pakistan border.

Human communities that inhabit the political borders live a transitional life, which is due to several socio-political and geo-climatic factors. The current cross-cultural ethnomedicinal study was conducted at the highly elevated Pakistan-India borders of the Western Himalayas in order to address the medicinal flora and folk knowledge of Balti and Shina communities. Ethnobotanical field surveys were conducted from May 2021 to September 2022. We used semi-structured interviews in order to collect the ethnomedicinal data. The collected data was analyzed using the relative frequency of the citations, use value, and Venn diagrams. A total of 140 interviews were conducted, which comprised of 90 (64.28%) Baltis and 50 (35.71%) Shinas. The interviews recorded 60 medicinal plant species that belong to 56 genera and 35 families. Asteraceae (5 spp.), Lamiaceae (5 spp.), and Apiaceae (4 spp.) were the families that were the most represented. These medicinally valued plants were being used for 55 health issues that are related to different body systems. Delphinium brunonianum, Thymus linearis, Hymenolaena candollei, Corydalis adiantifolia, and Seriphidium brevifolium were medicinal taxa with maximum RFCs and UV. The Baltis have comparatively more ethnomedicinal knowledge than the Shinas. The Baltis commonly used 22 (36.66%) species, which 34 (56.66%) were exclusive to the Baltis and 4 (6.66%) to the Shinas. Both ethnic groups collectively hold significant ethnobotanical knowledge that demands the preservation of risked folk knowledge, which is due to uncertain border situations, outmigration, and permeating allopathic drugs.

Connecting nutritional facts with the traditional ranking of ethnobotanically used fodder grasses by local farmers in Central Punjab of Pakistan.

The local farmers of Central Punjab, Pakistan have been using indigenous grasses as vital components of ruminant diets, but little is reported about their nutritional potential. Hence this study investigated nutritive potential of a selection of ethnobotanically important fodder grasses. Multiple nutritional parameters (proximate components, fibre fractions), secondary metabolites (phenolics, tannins) and in vitro digestibility values were determined. Furthermore, the legitimacy of ethnobotanical knowledge of local inhabitants about these grasses was also verified. The results suggested that majority (77%) of these grasses can be regarded as good quality fodders because of their high protein (169 g/kg) and good digestibility (457 g/kg) with moderate fibre (≤ 602 g/kg), lignin (≤ 50 g/kg) and secondary metabolites (total phenols ≤ 87 g/kg, total tannins ≤ 78 g/kg, condensed tannins ≤ 61 g/kg). Pearson correlation between nutritional parameters indicated that in vitro digestibility values were positively correlated with crude proteins (IVDMD, r = + 0.83 and IVOMD, r = + 0.83 respectively) and negatively correlated with fibre (NDF, r = - 0.91), ADF, r = - 0.84 and ADL, r = - 0.82) contents. Moreover, a positive relationship was identified between ethnobotanical knowledge and laboratory findings for studied grasses. Spearman correlation test showed that ranking of grasses based on ethnobotanical preferences were highly correlated (r values) with the laboratory results for CP (0.85), NDF (- 0.76), ADF (- 0.72) and ADL (- 0.62). The resilient complementarities between ethnobotanical preferences and nutritive analysis authenticate farmer's traditional knowledge, which needed to be aligned with the corresponding scientific data. Farmers can use these findings for appropriate fodder selection and development of precise supplements for feeding ruminants within a sustainable and economically viable livestock industry for food security.

Light and scanning electron microscopic comparative studies of geminivirus infected and healthy Eclipta alba (L.).

Eclipta alba (L.) is a valuable medicinal plant. However, its medicinal efficacy can be affected by geminivirus infection. Therefore, identification of healthy specimen is essential before to use as medicine. The present study provided the taxonomic characterization of geminivirus infected and healthy E. alba plant by studying apparent morphology and microscopic features through light and scanning electron microscopy. Before taxonomic characterization infected and healthy specimens were separated through molecular detection of geminivirus. Results of morphological studies reported that geminivirus infected E. alba plant showed systematic symptoms of infection like stunted growth, distortion and chlorosis of leaves, decrease in size of root, shoot and fruit, and so forth in comparison to healthy specimen. Anatomical findings reported that in both plants anomocytic and anisocytic types of stomata with multicellular warty trichomes were present. However, variations were observed in quantitative measures such as size of trichomes, epidermal, subsidiary and guard cells. Palynological observations identifies that both plants possessed tricolporate type of pollen but variation was mainly observed in size and shape of pollen, thickness of exine and intine, P/E ratio, pore size, interspecific difference, size of colpi, and pollen ornamentation. Overall this study concluded that both healthy and infected E. alba do not reported much variations in qualitative taxonomic features, but can be differentiated in terms of quantitative taxonomic evidences. Future studies are recommended for pharmacological analysis of both healthy and virus infected plants. RESEARCH HIGHLIGHTS: Eclipta alba has incredible therapeutic worth, but due to geminivirus infection the plant is affecting badly. Hence, the present studies give a comprehensive taxonomic report on the geminivirus infected and healthy plant species.

Indigenous knowledge and quantitative ethnobotany of the Tanawal area, Lesser Western Himalayas, Pakistan.

Ethnobotanical field surveys were carried out in the Tanawal area of the Lesser Himalayan Region, Khyber Pakhtunkhawa, Province from April 2016 to October 2017. The area is located between 34.36 (34° 21' 30 N) latitude and 73.07 (73° 4' 0 E) longitude with an average elevation of 1374 meters above sea level. Ethnomedicinal data were collected through Participatory Rural Appraisal (PRA), and participants were selected through the snow-boll technique. Semi-structured, in-depth and open-ended interviews were conducted. The data were quantitatively evaluated using ethnomedicinal indices i.e. Relative frequency of citation (RFCs), Fidelity level (FL), and Use Value (UV). The ethnobotanical data were also comparatively analyzed through the Jaccard Index (JI). The study yielded 66 medicinal plants in 62 genera and 43 families. Asteraceae and Solanaceae were the most important families with five medicinal taxa each. Regarding medicinal plant part utilization, leaves (43.28%) were used predominantly, followed by whole plant (14.92%) and fruits (14.92%). Decoction was the main drug formulation applied to 21 species (31.15%) and the oral route was most common (56.1%) while 31.2% of medicinal plants were used for both oral and topical applications. Fifty health disorders were recorded and grouped in 15 categories. Maximum species were used to treat gastrointestinal disorders i.e. 13 species, dermal problems (12 species), and respiratory tract ailments (9). The calculated RFCs ranged between 81 to 31. The most important medicinal plants were Acacia modesta, Citrullus vulgaris, Tamarindus indica, and Momordica charantia with an RGFC of 81 each. The UV ranged between 0.58 and 3.6. Medicinal taxa with the highest UV were Dioscorea deltoidea (3.6), Withania coagulans (3.3), Momordica charantia (3.5), Silybum marianum and Pyrus pashia (3.2). FL values showed that 28 (41.79%) species had a FL value below 50 (74.62%) while 39 (58.20%) had higher FL values. Momordica charantia, Tamarindus indica, Acacia modesta and Citrullus vulgaris were 95.2 each. The Jaccard Index (JI) values ranged from16.77 to 0.98. The current study also reported 16 medicinal plants, commonly used around the globe, have been rarely documented for their medicinal values in the local ethnomedicinal literature i.e. Althaea officinalis, Plantanus orientalis, Jasminum sombac, Maytenus royleana, Cucurbita maxima, Phyllanthus emblica, Citrullus vulgaris. Polygonatum verticilliatum, Caseria tomentosa, Cistanche tubulosa, Bambusa arundinacea, Schinus molle, Tamarindus indica, Pongamia pinnata, Citrus limon and Catharanthus roseus. However, 48 medicinal plants had been reported in the literature but the current study reported their novel medicinal uses. Important taxa should be established in botanical gardens for in-situ conservation, chemical investigation and sustainable utilization. It would also be effective to improve the livelihoods of the local population.

Morphological and palynological assessment of taxonomically problematic genus Paspalum based on light and scanning electron microscopy.

Present research work was carried out to clarify the variations among species of genus Paspalum morphologically and palynologically as this genus is taxonomically difficult due to having multiple similar morphologically overlapping characters which make it difficult to identify. Henceforth present research work was carried out to delimit taxa within the same genus by morphological and palynological tools through light microscopy (LM) and scanning electron microscopy (SEM). Both these tools are considered as the most useful taxonomic characters for taxonomically problematic genera. The results showed a lot of variations among morphological characters. In Paspalum dilatatum, the upper glume was ovate whereas in the other two species, the upper glume was elliptic. The upper glume apex found in P. dilatatum and Paspalum scrobiculatum was obtuse whereas in Paspalum distichum, upper glume apex was acute. Glume nerves showed variation in all three species. Paspalum distichum was 3 nerved, P. scrobiculatum was 5-7 nerved, and P. dilatatum was 5-9 nerved. All three species showed variation in lemma nerves. Paspalum scrobiculatum had 3 nerved lemma whereas in P. distichum 3-5 nerved and P. dilatatum 5-9 nerved lemma were present. In polar and equatorial view, pollen grains ranged from 25 (20-30) to 37.5 (30-45) µm. Paspalum distichum appeared to be the smallest in size whereas P. dilatatum was the largest. Exine thickness ranged from 0.75 (0.5-1) to 1.35 (1.2-1.5) µm. The higher value of pollen fertility was found in P. scrobiculatum as 87.69% and the lowest value was in P. distichum as 78.08%. Morphological keys were also given for correct identification.

Scanning electron microscopic imaging of palynological metaphors: A case study of family Poaceae taxa.

This study highlighted the taxonomic utilization of palynological metaphors for selected members (53) of family Poaceae. Multiple microscopic technique light and scanning electrons had been employed for detailed analysis. Results reported monad pollen type in all studied 53 members, which showed its limited taxonomic value up to family level. In relation to shape of pollen both polar and equatorial views strikingly differed from each other. Like semi angular pollen observed in Dactyloctenium aegyptium (L.) Wild. whereas round-minutely irregular pollen in Lolium temulentum L. Polar and equatorial diameter also showed variation, that is, Desmostachya bipinnata (L.) Stapf. can be differentiated from Sorghum bicolor (L.) Moench on diameter variation basis. A pore characteristic does not show much qualitative variation; however pore sizes differ species to species. The most frequently scarbate sculpturing was observed in 28 species followed by verrucate. Hence it can be said that pollen shape, polar, and equatorial diameters, pore size, P/E ratio, pore sculpturing are of good taxonomic value and holds a significant position in identification and delimitation of Poaceae taxa.

Natural Coumarins: Exploring the Pharmacological Complexity and Underlying Molecular Mechanisms.

Coumarins belong to the benzopyrone family commonly found in many medicinal plants. Natural coumarins demonstrated a wide spectrum of pharmacological activities, including anti-inflammatory, anticoagulant, anticancer, antibacterial, antimalarial, casein kinase-2 (CK2) inhibitory, antifungal, antiviral, Alzheimer's disease inhibition, neuroprotective, anticonvulsant, phytoalexins, ulcerogenic, and antihypertensive. There are very few studies on the bioavailability of coumarins; therefore, further investigations are necessitated to study the bioavailability of different coumarins which already showed good biological activities in previous studies. On the evidence of varied pharmacological properties, the present work presents an overall review of the derivation, availability, and biological capacities of coumarins with further consideration of the essential mode of their therapeutic actions. In conclusion, a wide variety of coumarins are available, and their pharmacological activities are of current interest thanks to their synthetic accessibility and riches in medicinal plants. Coumarins perform the valuable function as therapeutic agents in a range of medical fields.

Morphological and palynological assessment of some taxa of genus Echinochloa through light and scanning electron microscopy.

The present study was undertaken to gather information on selected species of Echinochloa based on their morphological and palyonological examination through light and scanning electron microscopy. As the Echinochloa species are most problematic grass weeds and exhibit high interspecific and intraspecific variability so due to their great morphological diversity it is difficult to recognize them. A lot of variation was observed in morphological features of studied taxa. In the present study, E. colona had more or less spreading dense racemes whereas in E. crus-galli racemes were linear to ovate. Upper glume surface was pubescent in E. colona, hispid in E. crus-galli and spinose in E. walteri. E. colona was comprised of five-nerved glumes whereas in E. crus-galli glumes were three-nerved. Although pollen basic characters in family Poaceae remains consistent but great variation was observed in pollen of studied taxa. The pollen size was quite variable among the species of genus Echinochloa. In polar view, the size of pollen grains ranged from 27.5 (20-35) µm to 40 (30-50) µm. E. crus-galli was appeared to be the smallest in size whereas E. colona was the largest.). P/E ratio ranged from 0.84 to 1.15 µm among the species. The highest value of pollen fertility was found in E. colona as 93.33% and lowest was in E. crus-galli as 87.50%. The morphological key of studied taxa is also given to study variations. In conclusion, it was observed that microscopy of morphological and palynological assessment can play a vital role in identification of taxonomically problematic taxa.

Application of multimicroscopic techniques (LM and SEM) in comparative palynological study of Asteroideae members, inhabited in Pakistan.

In the present study light and scanning electron microscopic approaches were used to study the palynological variations among 30 Asteroideae members. Variety of qualitative and quantitative palynological characters like pollen shape and type, pollen sculpturing, pollen size, P/E ratio, exine and intine thickness, interspecific difference, colpi size, pore size, spines length, spine width, and pollen fertility were observed. Results reported remarkable variations among pollen characters of studied taxa. The shape of pollen grains in polar and equatorial views varied from specie to specie such as spheroidal, prolate, oblate, subspheroidal, oblate spheroidal, prolate spheroidal, suboblate, and sub prolate. Bidens pilosa L. showed maximum pollen size in both polar and equatorial views, whereas minimum pollen size in polar view was found in Tetraneuris scaposa (DC.) Greene. (i.e., 30.5 µm) and in equatorial view was found in Cosmos sulphureus Cav. (25.5 µm).The highest P/E ratio (1.39 µm) was measured in Glebionis coronaria L. The values of exine and intine thickness also varied. Tricolporate, tetracolporate, trizonocolporate, pentoporate, and polypentoporate pollen types were examined. Spinateand echinate pollen sculpturingobserved under SEM. Variations in spine length and width also recorded. The maximum pollen fertility (98%) was measured in Chrysanthemum morifolium Ramat. and the lowest (56%) was recorded in Thymophylla tenuiloba (DC.) Small. It was concluded that the variations in qualitative and quantitative characters were seemed to be valuable for the taxonomic investigations of Asteroideae taxa.

Scanning Electron Microscopy-based palynological characterization of selected Euphorbiaceae taxa colonized in Pakistan.

The present study was carried out on scanning electron microscopic-based palynological characterization of selected 20 Euphorbiaceae taxa. In this study, important pollen qualitative and quantitative features of selected Euphorbiaceae species such as polar diameter, equatorial diameter, P/E ratio, sculpturing of pollen, exine thickness, intine thickness, presence or absence of colpi and length of colpi, and pollen fertility were examined. Remarkable variations in these pollen characters had been observed among these species. In both polar and equatorial views, Riccinus communis L. exhibited the highest pollen size (polar = 264.1 µm, Equatorial = 270 µm), while Acalypha wilkesiana Mull Arg. showed lowest pollen size (Polar = 17 µm, Equatorial = 18.5 µm). Most of the pollen varied from oblate to spheroidal in shape. With regard to the P/E ratio, Sapium sebiferum L. Roxb showed the highest ratio while Bischofia javanica Blume showed minimum, that is, 1.18 and 0.85, respectively. Intine and exine thickness also varied. Most of the pollen grains were monade. Tricolporate, tricolpate, monocolpate, pentaporate, and polyporate pollen aperture types have been observed. Most of the plants showed regular, reticulate, and echinate pollen sculpturing. Pollen fertility also varied from species to species but most of the plant showed pollen fertility between 70 and 80%. Through this work, it has been concluded that quantitative and qualitative pollen features are helpful at the specific level as well as the generic level and can provide a fruitful taxonomic solution.

Scanning electron microscopic screening; Can it be a taxonomic tool for identification of traditional therapeutic plants.

The limiting factor in the utilization of herbal medicine is the risk of replacement or substitution of their plant resources. Therefore, the authentication of medicinal plants before its use in herbal medicines is essentially required. Hence, the prime objective of this study was to provide some reliable morpho-anatomical and morpho-palynological tools for the identification of nine traditional therapeutic plants of district Lahore. For this study, scanning electron microscopic and light microscopic approaches has been employed. Results reported great variations in qualitative and quantitative morpho-anatomical features of the epidermal cell, stomatal complexes, trichomes, silica bodies and oil droplets. For example, Aegle marmelos L. can be distinguished from Cestrum diurnum L. based on epidermal cell shapes, as irregular cells in former and lobed cells shape in later one. Similarly unicellular, bicellular, and multicellular trichomes could be an identification basis for Cynodon dactylon L., Lantana camara L., and Pongamia pinnata L., respectively. In combination with these morpho-anatomical characters, SEM-based morpho-palynological characterization also reported diagnostic value. Both the Aegle marmelos and Lantana camara possessed similar cell shape (irregular) but can be delimited based on their pollen morphology as spheroidal pollen was reported in Aegle marmelos whereas prolate in Lantana camara. Moreover, quantitative characters also aid well for species delimitation, such as prolate pollen found in both Cestrum diurnum and Terminalia chebula Retz. but both of these varied in their P/E ratio, that is, 0.97 and 0.88, respectively. The leaf epidermal characters in correlation with morph-palynological traits showed potential for taxonomic resolution at the species level. RESEARCH HIGHLIGHTS: This study provides valuable morpho-anatomical and morpho-palynological tools for the nine traditional therapeutic plants of the district, Lahore, Pakistan. Light and scanning electron microscopic studies provided useful systematic data for the identification of studied plants. This systematic data include moprho-anatomical and morpho-palynological keys for the studied plants. This data can be used for future identification purposes.

Light and scanning electron microscopy-based foliar micro morphological tools for the identification of fodder grass taxa.

Fertile plain of Central Punjab Pakistan is rich with fodder grasses and from centuries the local inhabitants of this area have been using their regional grasses for ruminant feeding. However, they always faced difficulties in identification because of their overlapping vernacular names, more or less identical leaf shapes, indefinite variations in stem branching pattern, and reduced floral parts. Hence, the current study has provided a detailed and comprehensive micro-morphological analysis of 53 ethnobotanical fodder grass taxa. A variety of quantitative and qualitative leaf epidermal micromorphological traits was studied and results reported epidermal characters like stomatal index, silica bodies, prickles, microhairs, hook cells as most diagnostic in delimitation of species, and genera. As stomatal index was 79% in Poa annua while in its closely related species Poa infirmai was 85%. Similarly, Cenchrus ciliaris can be differentiated from Cenchrus pennisetiformis on the basis of silica body shape as butterfly shaped in former and dumbbell shaped in later one. Moreover, prickles were present in Chrysopogon aucheri while absent in Chrysopogon zizanioides. Hence, overall this study declared that diversity and variations in foliar micromorphological characters are valuable and supportive in the identification of grasses at the specific and generic level.

Implication of palynological techniques for the authentication of adulterated drugs traded with the same name in different herbal markets of district Lahore, Pakistan.

Adulteration in traded medicinal plants is a significant issue nowadays and use of these adulterated medicinal plants can impose harmful impact to end user. However, this problem can be overcome by ensuring the identification of traded medicinal plants which are used in making different herbal medicines. In this regard, palynological markers are considered to be an important taxonomic tool in the identification of original medicinal plant from its adulterant. Hence this study attempted to provide particular reliable palynological markers for distinguishing selected medicinal plants from their adulterants, that is, Cinnamomum verum versus Canella winterana, Cinnamomum tamala versus Cinnamomum obtusifolium, Gymnema sylvestre versus Gymnema lactiferum, Artemisia maritima versus Artemisia absinthium, Achillea millefolium versus Adhatoda vasaka, Sphaeranthus indicus versus Sphaeranthus africanus, Averrhoa carambola versus Butea monosperma, and Morus nigra versus Morus alba. Results demonstrated great variations in multiple palynological characters between original medicinal plant and its adulterant such as in pollen size, shape, colpi length, exine, intine thickness, and fertility. In equatorial view, circular to spheroidal shape of pollen was found in A. millefolium while oblate shape was observed in A. vasaka. Similarly B. monosperma pollen was 34 µm, whereas pollen of its adulterant A. carambola was 21 µm. Moreover, colpi length of A. maritima was 11.8 µm, whereas 4.5 µm in A. absinthium. Hence it can be concluded that palynological characters are commendably helpful in identification of genuine medicinal plant from its adulterant.

Ethnobotanical studies of fodder grass resources for ruminant animals, based on the traditional knowledge of indigenous communities in Central Punjab Pakistan.

BACKGROUND: Traditional knowledge of indigenous plants is pivotal in developing strategies to feed livestock sustainably in low input systems. Likewise, in Pakistan the indigenous people of Central Punjab have been using their regional grasses as a ruminant fodder for centuries. This study evaluated the indigenous traditional knowledge to ascertain the value of various fodder grasses to optimise their use to feed livestock in Central Punjab. METHODS: The snowball technique was employed to identify key informants who had relevant knowledge about different grasses in the study area. Semi-structured questionnaires, face-to-face interviews and site visits were used for describing the fodder grasses. The data were then analysed by using relative frequency citation and pairwise comparison methods to determine the order of priority among the listed fodder grasses. Furthermore, SPSS 22 software was used for descriptive statistics and interpretation of associations among studied parameters. Microsoft Excel was used to present data as % values and graphs. RESULTS: Overall, 53 grasses were described with ethnobotanical information regarding their uses for fodder, ethnoveterinary and other purposes. All these grasses belonged to the family Poaceae where the subfamily Panicoideae had the maximum number of 30 grasses. We categorized these grasses into high (A), medium (B) and low priority (C) groups where the group A grasses were reported as not only the most abundant but also the most palatable forages to all ruminants. Their higher demand was reflected by the feeding systems of both ad libitum grazing and feeding after cutting and mixing with other feeds. The study also revealed 37 previously unreported ethnoveterinary uses of these grasses. CONCLUSIONS: The results have reinforced the value of conserving ethnobotanical knowledge, being poorly documented previously, in developing strategies to feed livestock. It indicated the preferred fodder grasses as well as the possible reasons of their preference. The reported data need to be validated for nutritional and health benefits. This information could help the smallholder farmers in association with regional governments to propagate suitable fodder grasses for their use in sustainable livestock feeding to produce safe and healthy food for indigenous communities.