Traditional Wild Food Plants Gathered by Ethnic Groups Living in Semi-Arid Region of Punjab, Pakistan.

Wild edible food plants (WFPs) are valuable resources in the traditional food systems of many local cultures worldwide, particularly in underdeveloped regions. Understanding patterns of food preferences requires conducting cross-cultural food studies among various ethnic groups in a specific area. In this context, the current study aimed to record WFP use among five ethnic groups in Punjab, Pakistan, by interviewing 175 informants selected through snowball sampling. The indicator food species for different ethnic groups were calculated using indicator analysis based on the percentage of citations. A total of 71 wild food plants (WFPs) belonging to 57 genera and 27 families were observed in the study area. A high proportion of these wild food plants (WFPs) belonged to Fabaceae with eleven species (15%), followed by Moraceae with seven species (9%). Fruits were most widely used (43%), followed by leaves (19%), and shoots (16%). The majority (35 species, 49%) of plants of WFPs were eaten as cooked vegetables. A cross-cultural comparison revealed that four species overlapped among five ethnic groups (Arain, Jutt, Rajpot, Mewati, and Dogar). The Arain ethnic group gathered and consumed a remarkable number of wild plants (35 species), possibly due to a special connection with the general abundance of the local flora, and being close to nature by adopting professions more allied to WFPs in the study area. The analysis of indicator species revealed distinct significant indicator values (p ≤ 0.05) between the main food species among the various ethnic groups. Amaranthus viridis was a common indicator of food in all five ethnic groups, while Ziziphus nammularia was a common indicator food plant of the Mewati, Rajpot, and Jutt ethnic groups; these plants are important in local diets, especially during times of food scarcity brought on by disease or drought. In addition, the current study reports 20 WFPs that have been rarely documented as human food in Pakistan's ethnobotanical literature. Future development plans should consider biocultural heritage and pay appropriate attention to local ecological knowledge, dynamics, and historical exchanges of traditional food systems.

Phytochemical Screening, Antioxidant and Antifungal Activities of Aconitum chasmanthum Stapf ex Holmes Wild Rhizome Extracts.

Aconitum chasmanthum Stapf ex Holmes, an essential and critically endangered medicinal plant from Kashmir Himalayas, was studied for its antioxidant and antifungal properties. The shade-dried powdered rhizome was extracted sequentially with hexane, ethyl acetate, and methanol. These subsequent fractions were evaluated for total phenolic content (TPC); total flavonoid content (TFC); antioxidant assays, such as 1,1-diphenyl 1-2-picryl-hydrazyl (DPPH); ferric-reducing antioxidant power (FRAP); superoxide radical scavenging (SOR); hydroxyl radical scavenging (OH) and antifungal activity using the poisoned food technique. Highest TPC (5.26 ± 0.01 mg/g) and TFC (2.92 ± 0.04 mg/g) were reported from methanolic extracts. The highest values of radical scavenging activities were also observed in methanolic extracts with IC50 values of 163.71 ± 2.69 µg/mL in DPPH, 173.69 ± 4.91 µg/mL in SOR and 159.64 ± 2.43 µg/mL in OH. The chemical profile of ethyl acetate extract was tested using HR-LCMS. Methanolic extracts also showed a promising inhibition against Aspergillus niger (66.18 ± 1.03), Aspergillus flavus (78.91 ± 1.19) and Penicillium notatum (83.14 ± 0.97) at a 15% culture filtrate concentration with minimum inhibitory concentration (MIC) values of 230 µg/mL, 200 µg/mL and 190 µg/mL, respectively. Overall, the methanolic fractions showed significant biological potential, and its pure isolates might be used to construct a potential new medicinal source.

Alien woody plants are more versatile than native, but both share similar therapeutic redundancy in South Africa.

Understanding why alien plant species are incorporated into the medicinal flora in several local communities is central to invasion biology and ethnobiology. Theories suggest that alien plants are incorporated in local pharmacopoeias because they are more versatile or contribute unique secondary chemistry which make them less therapeutically redundant, or simply because they are locally more abundant than native species. However, a lack of a comprehensive test of these hypotheses limits our understanding of the dynamics of plants knowledge, use and potential implications for invasion. Here, we tested the predictions of several of these hypotheses using a unique dataset on the woody medicinal flora of southern Africa. We found that the size of a plant family predicts the number of medicinal plants in that family, a support for the non-random hypothesis of medicinal plant selection. However, we found no support for the diversification hypothesis: i) both alien and native plants were used in the treatment of similar diseases; ii) significantly more native species than alien contribute to disease treatments particularly of parasitic infections and obstetric-gynecological diseases, and iii) alien and native species share similar therapeutic redundancy. However, we found support for the versatility hypothesis, i.e., alien plants were more versatile than natives. These findings imply that, although alien plant species are not therapeutically unique, they do provide more uses than native plants (versatility), thus suggesting that they may not have been introduced primarily for therapeutic reasons. We call for similar studies to be carried out on alien herbaceous plants for a broader understanding of the integration of alien plants into the pharmacopoeias of the receiving communities.

Methodological advances for hypothesis-driven ethnobiology.

Ethnobiology as a discipline has evolved increasingly to embrace theory-inspired and hypothesis-driven approaches to study why and how local people choose plants and animals they interact with and use for their livelihood. However, testing complex hypotheses or a network of ethnobiological hypotheses is challenging, particularly for data sets with non-independent observations due to species phylogenetic relatedness or socio-relational links between participants. Further, to account fully for the dynamics of local ecological knowledge, it is important to include the spatially explicit distribution of knowledge, changes in knowledge, and knowledge transmission and use. To promote the use of advanced statistical modelling approaches that address these limitations, we synthesize methodological advances for hypothesis-driven research in ethnobiology while highlighting the need for more figures than tables and more tables than text in ethnobiological literature. We present the ethnobiological motivations for conducting generalized linear mixed-effect modelling, structural equation modelling, phylogenetic generalized least squares, social network analysis, species distribution modelling, and predictive modelling. For each element of the proposed ethnobiologists quantitative toolbox, we present practical applications along with scripts for a widespread implementation. Because these statistical modelling approaches are rarely taught in most ethnobiological programs but are essential for careers in academia or industry, it is critical to promote workshops and short courses focused on these advanced methods. By embracing these quantitative modelling techniques without sacrificing qualitative approaches which provide essential context, ethnobiology will progress further towards an expansive interaction with other disciplines.

Effective antioxidant, antimicrobial and anticancer activities of essential oils of horticultural aromatic crops in northern Egypt.

BACKGROUND: Identifying ornamental plants as new natural antioxidant and antimicrobial sources is always of great importance for the ornamental and horticultural industries. METHODS: The antimicrobial activities of leaves and fruits peel essential oils of twelve ornamental and horticultural crops were determined by screening against wide spectrum of fungi and bacteria, and their respective in vitro antioxidant capacity was evaluated. Furthermore, the anticancer activities against several cancer cells, and one normal human cell line (HEK-293) were examined. RESULTS: Origanum vulgare L. essential oil showed the best antioxidant, antibacterial and anticancer activities compared to screened crops by means of the DPPH and linoleic acid assays for antioxidants, MIC and MBC values for antibacterial activities and IC50 for antiproliferative activities. Such important activities in O. vulgare was attributed to high pulegone ratio (77.45%) as revealed by the GC/MS assay. Rosmarinus officinallis L. essential oil showed the highest antifungal activities by means of lowest MIC and MFC values which might be attributed to 1, 8-cineole (19.60%), camphor (17.01%) and α-pinene (15.12%). CONCLUSION: We suggest that oxygenated monoterpenes (i.e. linalool, terpinen-4-ol and pulegone) and monoterpene hydrocarbons play an important role in the essential oil antioxidant, antibacterial, antifungal and anticancer activities of diverse Egyptian ornamental and horticultural crops. Some species showed bioactivities similar to standards compounds and might be suitable for pharmaceutical and food industries.

In vitro antibacterial, antifungal and antioxidant activities of Eucalyptus spp. leaf extracts related to phenolic composition.

The crude methanolic extracts from leaves of Eucalyptus camaldulensis L., E. camaldulensis var obtusa and E. gomphocephala grown in Egypt were investigated to explore their chemical composition as well as their antibacterial, antifungal and antioxidant activities. Major phenolics found were ellagic acid, quercetin 3-O-rhamnoside, quercetin 3-O-b-D-glucuronide, caffeic acid and chlorogenic acid. The antioxidant activities were examined by the 2,2'-diphenypicrylhydrazyl (DPPH) and ß-Carotene-linoleic acid assays. E. camaldulensis extracts showed the highest phenolic content, antioxidant and antimicrobial activities compared to other cultivars. MIC values reported for antibacterial activity of E. camaldulensis ranged from 0.08 µg/mL (Bacillus cereus) to 0.22 µg/mL (Staphylococcus aureus), while MBC values ranged from 0.16 µg/mL (Dickeya solani and B. cereus) to 0.40 µg/mL (S. aureus). The inhibitory activities against growth of bacteria and fungi used is an indication that E. camaldulensis a might be useful resource for the development and formulation of antibacterial and antifungal drugs.

The first initiative of DNA barcoding of ornamental plants from Egypt and potential applications in horticulture industry.

DNA barcoding relies on short and standardized gene regions to identify species. The agricultural and horticultural applications of barcoding such as for marketplace regulation and copyright protection remain poorly explored. This study examines the effectiveness of the standard plant barcode markers (matK and rbcL) for the identification of plant species in private and public nurseries in northern Egypt. These two markers were sequenced from 225 specimens of 161 species and 62 plant families of horticultural importance. The sequence recovery was similar for rbcL (96.4%) and matK (84%), but the number of specimens assigned correctly to the respective genera and species was lower for rbcL (75% and 29%) than matK (85% and 40%). The combination of rbcL and matK brought the number of correct generic and species assignments to 83.4% and 40%, respectively. Individually, the efficiency of both markers varied among different plant families; for example, all palm specimens (Arecaceae) were correctly assigned to species while only one individual of Asteraceae was correctly assigned to species. Further, barcodes reliably assigned ornamental horticultural and medicinal plants correctly to genus while they showed a lower or no success in assigning these plants to species and cultivars. For future, we recommend the combination of a complementary barcode (e.g. ITS or trnH-psbA) with rbcL + matK to increase the performance of taxa identification. By aiding species identification of horticultural crops and ornamental palms, the analysis of the barcode regions will have large impact on horticultural industry.

Enhancement of Calibrachoa growth, secondary metabolites and bioactivity using seaweed extracts.

BACKGROUND: Calibrachoa x hybrida (Solanaceae) cultivars are widely used in North and South America as ornamental plants. Their potential as a source of antimicrobial compounds might be enhanced by seaweed extract (SWE) applications. METHODS: SWE of Ascophyllum nodosum were applied at 5 and 7 ml/L as a soil drench or foliar spray on Calibrachoa cultivars of Superbells® 'Dreamsicle' (CHSD) and Superbells® 'Frost Fire' (CHSF). The total phenolics, tannins and antioxidants composition as well as specific flavonols in leaf extracts were determined. Further, the chemical composition of SWE was assessed. RESULTS: The drench and foliar SWE treatments significantly enhanced Calibrachoa cultivars leaf number and area, dry weight, plant height, antioxidant capacity as well as phenolic, flavonols and tannin content. The increased growth and composition of phenols, flavonols and tannins was attributed to the stimulatory effects of SWE mineral composition. The antifungal activity of Calibrachoa cultivars was significantly enhanced following SWE treatments and the minimum inhibitory concentration (MIC) and minimum fungicidal concentration (MFC) were in the range of 0.07-0.31 mg/ml and from 0.16 to 0.56 mg/ml, respectively. Moreover, antibacterial activity was significantly increased and the MIC and minimum bactericidal concentration (MBC) measurements were in the range of 0.06-0.23 mg/ml and from 0.10 to 0.44 mg/ml, respectively. The most sensitive fungus to SWE treatments was C. albicans and the most sensitive bacterium was E. cloacae. CONCLUSION: The results suggest that enhanced antifungal and antibacterial activities might be attributed to significant increases of phenolic, flavonols and tannin contents, which ultimately enhance the potential of Calibrachoa as a natural source of alternative antibiotics.

In vitro antioxidant, antifungal and antibacterial activities of five international Calibrachoa cultivars.

The total phenolic, flavonoid and tannin contents in leaf extracts of Calibrachoa x hybrida (C.h.) (Solanaceae) international cultivars, as well as their overall antioxidant activities using DPPH and linoleic acid assays, were investigated. Furthermore, the antifungal and the antibacterial activities were examined against a wide spectrum of micro-organisms. DPPH and linoleic acid assays ranged from 62.1 to 80.1% and of 74.1-93.4%, respectively. C.h. Superbells® Trailing Rose (CHST), C.h. Superbells® Frost Fire, C.h. Superbells® Strawberry Punch, C.h. Superbells® Dreamsicle and C.h. Superbells® Plum (CHSP) varied in their antifungal and the antibacterial activities against a wide spectrum of micro-organisms. CHSP exhibited the highest antioxidant, antifungal and antibacterial activities followed by CHST. These activities might be attributed to the presence of phenolic, flavonoid and tannin compounds, indicating that these cultivars might be potential sources of therapeutic substances.

In vitro Antioxidant and Antimicrobial Effects of Ceratostigma plumbaginoides.

Bioactive compounds, including phenols, flavonoids, and tannins, were quantified in leaves, stems and roots of methanol, n-butanol, diethyl ether and n- hexane extracts of Ceratostigina plumbaginoides Bunge. (Plumbaginaceae) ornamental plants. The antioxidant capacity was measured by the DPPH and linoleic acid assays. The total bioactive compounds, as well as the antioxidant capacities, were the highest in the leaves compared with stems and roots. The -methanolic, n-butanol, diethyl ether and n-hexane leaf extracts varied in their antibacterial and antifungal activities. In general, the most sensitive bacterium to leaf extracts was Bacillus cereus and the most resistant was Staphyllococcus aureus, while the most sensitive fungus was Aspergillusflavus and the most resistant one was Penicillium ochrochloron. As the methanolic leaf extract was the most active, it was subjected to column chromatography and two compounds were isolated and identified as 1 (5-hydroxy-2-methyl-1,4-naphthoquinone / plumbagin) and 2 (3,3'-biplumbagin). Compounds 1 and 2 showed the highest antibacterial and antifungal activities compared with other extracts tested. The MIC and MBC values for the most active compound 1 were in the range of 0.001 - 0.09 and 0.004 - 0.21 mg mL⁻¹, while MIC and MFC were determined as 0.001 - 0.11 and 0.002 - 0.19 mg mL⁻¹, respectively. The isolated compounds and leaf extracts showed also equal or higher antimicrobial activities compared with antibiotics/commercial reagents which indicate that the plant might be useful for drug development. This is the first report on the antibacterial and antifungal activities, as well as the antioxidant properties of the tested plant parts and isolated compounds.

Phylogenetic exploration of commonly used medicinal plants in South Africa.

The rapid growth rate of human population, along with the public health crisis encountered in many regions, particularly in developing world, creates an urgent need for the discovery of alternative drugs. Because medicinal plants are not distributed randomly across lineages, it has been suggested that phylogeny along with traditional knowledge of plant uses can guide the identification of new medicinally useful plants. In this study, we combined different statistical approaches to test for phylogenetic signal in 33 categories of plant uses in South Africa. Depending on the null models considered, we found evidence for signal in up to 45% of plant use categories, indicating the need for multiple tests combination to maximize the chance of discovering new medicinal plants when applying a phylogenetic comparative approach. Furthermore, although there was no signal in the diversity of medicinal uses-that is, total number of medicinal uses recorded for each plant-our results indicate that taxa that are evolutionarily closely related have significantly more uses than those that are evolutionarily isolated. Our study therefore provides additional support to the body of the literature that advocates for the inclusion of phylogeny in bioscreening medicinal flora for the discovery of alternative medicines.

Efficacy of the core DNA barcodes in identifying processed and poorly conserved plant materials commonly used in South African traditional medicine.

Medicinal plants cover a broad range of taxa, which may be phylogenetically less related but morphologically very similar. Such morphological similarity between species may lead to misidentification and inappropriate use. Also the substitution of a medicinal plant by a cheaper alternative (e.g. other non-medicinal plant species), either due to misidentification, or deliberately to cheat consumers, is an issue of growing concern. In this study, we used DNA barcoding to identify commonly used medicinal plants in South Africa. Using the core plant barcodes, matK and rbcLa, obtained from processed and poorly conserved materials sold at the muthi traditional medicine market, we tested efficacy of the barcodes in species discrimination. Based on genetic divergence, PCR amplification efficiency and BLAST algorithm, we revealed varied discriminatory potentials for the DNA barcodes. In general, the barcodes exhibited high discriminatory power, indicating their effectiveness in verifying the identity of the most common plant species traded in South African medicinal markets. BLAST algorithm successfully matched 61% of the queries against a reference database, suggesting that most of the information supplied by sellers at traditional medicinal markets in South Africa is correct. Our findings reinforce the utility of DNA barcoding technique in limiting false identification that can harm public health.