Biodiversity and Environmental Research Center

The incidence of Tomato brown rugose fruit virus (ToBRFV) and biological and molecular characterization of the Palestinian isolates of ToBRFV are described in this study. Symptomatic leaf samples obtained from Solanum lycopersicum L. (tomatoes) and Nicotiana tabacum L. (cultivated tobacco) plants were tested for tobamoviruses infection by reverse transcription polymerase chain reaction. Tomato leaf samples collected from Tulkarm and Qalqilia are infected with ToBRFV-PAL with an infection rate of 76% and 72.5%, respectively. Leaf samples collected from Jenin and Nablus were found to be mixed infected with ToBRFV-PAL and Tobacco mosaic virus (TMV) (100%). Sequence analysis of the ToBRFV-PAL genome showed that the net average nucleotide divergence between ToBRFV/F48-PAL strain and the Israeli and Turkish strains was 0.0026398±0.0006638 (±standard error of mean), while it was 0.0033066±0.0007433 between ToBRFV/F42-PAL and these two isolates. In the phylogenetic tree constructed with the complete genomic sequence, all the ToBRFV isolates were clustered together and formed a sister branch with the TMV. The sequenced Palestinian isolates of ToBRFV-PAL shared the highest nucleotide identity with the Israeli ToBRFV isolate suggesting that the virus was introduced to Palestine from Israel. The findings of this study enhance our understanding of the biological and molecular characteristics of ToBRFV which would help in the management of the disease.

This study aims at providing an updated checklist of the native vascular flora of the Palestinian West Bank and Gaza Strip (State of Palestine, SP), serving as a taxonomic and nomenclatural basis for botanical research and encouraging new floristic surveys and biosystematic studies. The study provides an up-to-date checklist of native vascular taxa of the SP and their distribution within the plant districts in the country. This is the very first annotated checklist for the native vascular plants of the SP which incorporates recent name changes, new distribution records, habitat, herbarium specimens catalouge, Red List criteria, wild edible plants, endemism and use in Traditional Palestinian Herbal Medicine. The quantitative analysis of the flora has shown that the SP hosts 1826 taxa, distributed in 686 genera and 108 families; five taxa are gymnosperms, nine taxa are Pteridophytes and 1812 taxa are angiosperms. The most represented families are Leguminosae (222 taxa, 12.2%), Asteraceae (197, 10.2%) and Poaceae (196, 10.7%), while the most represented genera are Trifolium (38, Leguminosae), Silene (32, Caryophyllaceae), Astragalus (27, Leguminosae), Medicago (26, Leguminosae), Allium (25, Amaryllidaceae) and Euphorbia (25, Euphorbiaceae). Annuals (52.4%), Hemicryptophytes (20.2%) and Chamaephytes (12.2%) are the most represented life-forms amongst the SP flora. The richest plant districts in the SP vascular plant taxa are Gaza Strip (GS) (1216 taxa), Jerusalem and Hebron Mountains (JHM) (1235) and Nablus Mountains (NM) (1126). Agglomerative hierarchical clustering (AHC) permitted the division of SP into two main regions, based on the existence of vascular plant taxa: Region 1 (western plant districts with 1128-1237 taxa) with higher water availability and temperate Mediterranean climate which permit the establishment of more than 65% of the total SP flora in these districts and Region 2 (eastern plant districts with 571-698 taxa), characterised by desert and semi-desert conditions, as well as the presence of alluvial and co-alluvial soils, which allow the survival of lower numbers of plant taxa. One hundred and sixty-five taxa of the SP flora are endemic and near-endemic. However, in comparison with some countries of the Mediterranean Basin, this number is below the average endemism concentration, along with other southern arid countries, such as Tunisia and Egypt. In total, there are 102 threatened plant taxa, belonging to 39 families and 83 genera representing 5.6% of the total plants in the SP. IUCN and the Conservation Measures Partnership (CMP) unified classification of direct threats for SP Red-Listed plants has shown a high extinction risk to the Palestinian threatened wild flora, with 76.5% of the threatened species being either critically endangered (CR) or endangered (EN); only 23.5% were vulnerable (VU). However, several taxa are threatened by numerous factors including small population size, human activities, for example, conversion of traditional to intensive agriculture accompanied by deep ploughing and the application of pesticides, urban development and construction, global climatic change, drying of marshes and wetlands, quarrying, fires and pollution. This checklist can help focus conservation efforts and provide a framework for research, protection and policy applications for the SP flora, especially for the endemic and threatened plants.

Plumbago europaea L. is a plant utilized in Palestinian ethnomedicine for the treatment of various dermatological diseases. The current investigation was designed to isolate plumbagin from P. europaea leaves, roots and for the first time from the stems. Moreover, it aimed to evaluate the antimycotic activity against three human fungal pathogens causing dermatophytosis, also against an animal fungal pathogen. The qualitative analysis of plumbagin from the leaves, stems, and roots was conducted using HPLC and spectrophotometer techniques, while the structure of plumbagin was established utilizing Proton and Carbon-13 Nuclear Magnetic Resonance (NMR) and Infrared (IR) techniques. The entire plant constituents were determined by GC-MS. Moreover, the antimycotic activity against Ascosphaera apis, Microsporum canis, Trichophyton rubrum, and Trichophyton mentagrophytes was assessed utilizing the poison food technique method. The percentage of plumbagin recorded in the leaves, stems, and roots was found to be 0.51±0.001%, 0.16±0.001%, and 1.65±0.015%, respectively. The GC-MS examination declared the presence of 59 molecules in the plant extract. The plant extract and pure plumbagin exhibited complete inhibition against all tested dermatophytes at 6.0mg/mL for the extracts and 0.2mg/mL for plumbagin. P. europaea root is the best source of plumbagin and the plant extract could represent a potential drug candidate for the treatment of dermatophytosis infections. Further studies required to design suitable dosage forms from the natural P. europaea root extracts or plumbagin alone, to be utilized for the treatment of dermatological and veterinary ailments.

Fig mosaic is a viral disease (FMD) that spreads in Palestinian common fig (Ficus carica L.) orchards. Recognizing the economic value of fig plants and the harmful nature of FMD, the disease poses a significant threat to the economy of the fig production in Palestine. We applied the reverse transcription and amplification (RT-PCR) and PCR technique to leaf samples of 77 trees and 14 seedlings of 17 fig cultivars. The samples were collected from orchards in the main fig-growing provinces of the Palestinian West Bank, to assess the prevalence of viruses associated with FMD, and confirm a possible link of symptoms with viruses detected. Four viruses were detected: Fig mosaic virus (FMV), Fig badnavirus-1 (FBV-1), Fig leaf mottle-associated virus 2 (FLMaV-2), and Fig fleck-associated virus (FFkaV). FMV and FBV-1 were found in all tested fig plants (100%), while FLMaV-2 and FFkaV were detected in 61.5% and 33% of the fig samples, respectively. The high incidence of FBV-1 in the newly propagated symptomatic and symptomless seedlings from different cultivars may be an indication that FBV-1 is integrated into the genome of the fig in a cultivar nondis-criminatory manner. Very weak or no association was detected between FMD symptoms severity in the 17 Palestinian fig cultivars with the various viruses' combinations observed (i.e., number of the viruses infecting the plant). These results support the notion that FMD symptom severity expression is likely to be controlled by a combination of FMV infection, cultivars, and environmental factors, rather than the number of viruses infecting the plant.

Lavandula pubescens Decne (LP) is one of the three Lavandula species growing wildly in the Dead Sea Valley, Palestine. The products derived from the plant, including the essential oil (EO), have been used in Traditional Arabic Palestinian Herbal Medicine (TAPHM) for centuries as therapeutic agents. The EO is traditionally believed to have sedative, anti-inflammatory, antiseptic, antidepressive, antiamnesia, and antiobesity properties. This study was therefore aimed to assess the in vitro bioactivities associated with the LP EO. The EO was separated by hydrodistillation from the aerial parts of LP plants and analyzed for its antioxidant, antimicrobial, anticholinesterase, and antilipase activities. GC-MS was used for phytochemical analysis. The chemical analysis of the EO composition revealed 25 constituents, of which carvacrol (65.27%) was the most abundant. EO exhibited strong antioxidant (IC 50 0.16–0.18 μ L/mL), antiacetylcholinesterase (IC 50 0.9 μ L/mL), antibutyrylcholinesterase (IC 50 6.82 μ L/mL), and antilipase (IC 50 1.08 μ L/mL) effects. The EO also demonstrated high antibacterial activity with the highest susceptibility observed for Staphylococcus aureus with 95.7% inhibition. The EO was shown to exhibit strong inhibitory activity against Candida albicans (MIC 0.47 μ L/mL). The EO was also shown to possess strong antidermatophyte activity against Microsporum canis , Trichophyton rubrum , Trichophyton mentagrophytes, and Epidermophyton floccosum (EC 50 0.05–0.06 μ L/mL). The high antioxidant, enzyme inhibitory, and antimicrobial potentials of the EO can, therefore, be correlated with its high content of monoterpenes, especially carvacrol, as shown by its comparable bioactivities indicators results. This study provided new insights into the composition and bioactivities of LP EO. Our finding revealed evidence that LP EO makes a valuable natural source of bioactive molecules showing substantial potential as antioxidant, neuroprotective, antihyperlipidemic, and antimicrobial agents. This study demonstrates, for the first time, that LP EO might be useful for further investigation aiming at integrative CAM and clinical applications in the management of dermatophytosis, Alzheimer’s disease, and obesity.

In the present study, the medicinal aromatic plant Mentha spicata has been investigated as a source of essential oil (EO) and pharmaceuticals. The quantity and composition of EO from M. spicata cultivated in Palestine were analyzed seasonally over a three-year period. A significantly higher EO content was produced in summer and fall months (2.54–2.79%). Chemical analysis of EO revealed 31 compounds with oxygenated monoterpenes (90%) as the most abundant components followed by sesquiterpene and monoterpene hydrocarbons (6 and 3%, respectively). M spicata can be characterized as a carvone chemotype (65%). EO and carvone have shown strong inhibitory activities against the principal enzymes associated with Alzheimer’s disease (AD) and overweight diseases (cholinesterase and porcine pancreatic lipase) and also shown strong antidermatophytic activity against Microsporum canis , Trichophyton rubrum , T. mentagrophytes , and Epidermophyton floccosum . The pancreatic lipase inhibition and the synergism showed the potential activity of M. spicata EO and carvone and that their combinations with standard drugs can be useful for the treatment of obesity and overweight. The results also demonstrated that, in addition to their significant inhibitory activity against biofilm formation of methicillin-resistant Staphylococcus aureus (MRSA), M. spicata EO and carvone had a strong inhibitory effect on metabolic activity and biomass of the preformed biofilm. The current study supports the utilization of M. spicata EO as a traditional medicine and opens perceptions to find more potent substances in the EO for the management of obesity, AD, and dermatophytosis and for combating drug-resistant bacterial infections.

Abstract: Background: Crop landraces embody a source of beneficial genes potentially providing endurance to environmental stress and other agronomic qualities including yield. Our study included eighty-eight snake melon accessions (Cucumis melo var. flexuosus) collected from 9 districts in the Palestinian West-Bank. These accessions represent four landraces of Palestinian snake melon: Green, and White Baladi, and Green, and White Sahouri. Results: This is the first report on successful application of genotyping by sequencing in snake melon. 9750 single-nucleotide polymorphism (SNP) and 7400 DArTseq genetic markers were employed to evaluate genetic biodiversity and population structure of Palestinian snake melon germplasm collection. Clustering based on neighbor-joining-analysis, principle coordinate and Bayesian model implemented in Structure showed that patterns of genetic diversity of snake melon landraces depends on their geographical source and unraveled the presence of two major local landraces (Sahouri, and Baladi) with accessions from each group clustering together. A significant correlation was observed between both types of markers in Mantel correlation test. A significant association between genetic and geographic matrices (P<0.0001) was also detected. AMOVA indicated that majority of variation (90%) was due to the difference within accessions. Conclusion: The Palestinian landraces seem to have unique genes that may allow the enhancement of the global snake melon gene pool and developments of the plant production worldwide. Our subsequent objective is to detect genotypes with promising qualities and to conduct association mapping studies concentrating on Fusarium-wilt resistance, yield, and environmental stresses. Keywords: Palestinian snake melon landraces, Genetic diversity, Cucumis melo var. flexuosus, Population structure, SNP, genotyping by sequencing, DArT-seq.

We evaluated the antioxidant and porcine pancreatic lipase inhibition (PPLI) activities of 90 plants extract. The antioxidant activity was measured using the free-radical scavenging capacity (DPPH), and reducing power (RP) assays. The pancreatic lipase inhibition assay was used to determine the PPLI activity of plant extracts. Among the 90 plant extracts examined, 41.0 % crude extracts showed anti-lipase activity of more than 50%. The most active plants by means of IC50 value were: Camellia sinensis (0. 5 mg/ml), Ceratonia siliqua (leaves) (0.8 mg/mL), Curcuma longa (0.8 mg/mL), Sarcopoterium spinosum (1.2 mg/mL), and Mentha spicata (1.2 mg/mL). The antioxidant activity of plant extracts using the DPPH and RP assays reveals comparable results. The most active antioxidant extracts using both assays were the leaves and fruit epicarp of Rhus coriaria, areal parts of Sarcopoterium spinosum, and leaves of Ceratonia siliqua. Our results suggest natural resources that possess strong antioxidant and pancreatic lipase inhibitory activities with potential applications in the treatment and prevention of obesity and overweight. The extracts of Camellia sinensis, Ceratonia siliqua, Curcuma longa, Sarcopoterium spinosum, and Mentha spicata were proved to have a great potential as antioxidants and anti-obesity agents. Keywords: Antiobesity, orlistat, weight-reducing effects, pancreatic lipase inhibition

ABSTRACT Environmental conditions, including irrigation water quality, are known to affect secondary metabolite production in plants and hence the composition and activity of essential oils of aromatic plants. The aim of this study is to evaluate the effect of irrigation with secondary-treated effluent, as compared with potable water irrigation, on the composition, biological activities and yield of essential oil (EO) from Origanum syriacum L. var. syriacum (OSS). The EO from OSS cultivated in Palestine under secondary treated municipal effluent compared with potable water irrigation were analyzed seasonally over a two-year period for oil quantity and composition, antioxidant activities 1,1- diphenyl-2-picrylhydrazyl (DPPH), 2,2 azino-bis (3-ethylbenzothiazloine-6-sulfonic acid) radical cation (ABTS), ferric reducing antioxidant power (FRAP), antimicrobial and inhibitory properties against acetylcholinesterase (AChE), butyrylcholinesterase (BChE), and porcine pancreatic lipase (PPL). The quality of the irrigation water did not affect the essential oil yield quantity and quality demonstrating the potential of this marginal water source for agri-industrial production of OSS. Under both irrigation water qualities, the EO contained 35 constituents, of which thymol (40.4–46.7%), and carvacrol (35.1–41.8%) were the most abundant. EOs from plants of both irrigation water qualities exhibited a strong free radical scavenging ability and reducing power activity, and high AChE, BChE, and PPL inhibitory activities. The EOs also showed a moderate antibacterial activity against most test bacterial strains, with high antibacterial activity against Staphylococcus aureus with 117 percent inhibition compared with chloramphenicol (30 μg/disc). The high inhibitory activity of the EOs on AChE, BChE, and PPL, and its high antioxidant and antibacterial activities may be due to high concentrations of monoterpenes, especially thymol and carvacrol. The data demonstrate that secondary-treated municipal effluents can be successfully utilized for irrigation of industrial crop production setups for essential oil production from OSS (as a source of natural antioxidants, anticholinesterase, antiobesity and antistaphylococcal agents) without compromising yield quantity and bioactivities compared with potable water irrigation. You can download at: https://authors.elsevier.com/a/1WCVt3IipncTlk

Background Gastrointestinal parasites are one of the main restrictions to small ruminant production. Their pathological importance is primarily related to the major production losses, in quantity or quality, induced by the direct action of worms. Control of these parasites is based exclusively on the frequent use of anthelmintic drugs. However, the resistance to anthelmintics in worm populations after commercialisation of chemical drugs is now widespread. Therefore, there is a need to find new natural resources to ensure sustainable and effective treatment and control of these parasites. The aim of this study was to evaluate the anthelmintic activity, as minimum inhibitory concentration (IC50 mg/mL), of different plant extracts using larval exsheathment inhibition assay using a two-species but steady population of parasitic nematodes (ca. 20% Teladorsagia circumcinta and 80% Trichostrongylus colubriformis). Results The study showed that the ethanolic extracts of 22 out of the 48 plant extracts, obtained from 46 plant species, have an inhibitory effect >50% (at concentrations of 100 mg/mL) on the third stage larvae (L3) of the nematodes exhibited the strongest inhibition activity (94%) with IC50 of 0.02 mg/mL, where other members of the Rhamnaceae family have shown to possess strong anthelmintic activity (70–89%). Conclusions Plant extracts are potential rich resources of anthelmintics to combat helminthic diseases. Our results suggest that extracts from Rhamnus elaternus, Epilobium hirsutum, Leucaena leucocephala and Rhamnus palaestinus have promising anthelmintic activity, with potential applications in animal therapeutics and feed.

Genetic diversity in 50 snake melon accessions collected from Palestine (West Bank) was assessed by examining variation in 17 phenotypic characters. These accessions belonged to four important landraces of Cucumis melo var. flexuosus: green Baladi (GB), white Baladi (WB), green Sahouri (GS), and white Sahouri (WS). Principal component analysis (PCA) and a dendrogram were performed to determine relationships among populations and to obtain information on the usefulness of those characters for the definition of cultivars. PCA revealed that secondary fruit skin color, flesh color, primary fruit skin color, and secondary skin color pattern were the principal characters to discriminate melon accessions examined in the present study. According to the scatter diagram and dendrogram, landraces of Cucumis melo var. flexuosus: GB, WB, GS, and WS formed different clusters. However, based on Euclidean genetic coefficient distances, GB and WB had the least degree of relatedness with GS and WS, indicating distantly related landraces (Baladi and Sahouri). On the other hand, the highest degree of relatedness was detected between WS on one hand, and both GS and WB on the other indicating closely related cultivars. Fruit traits variability among the different snake melon landraces was evaluated and discussed in this study. This evaluation of fruit trait variability can assist geneticists and breeders to identify populations with desirable characteristics for inclusion in cultivars breeding programs. Key words: Cucumis melo var. flexuosus, Snake melon, phenotypic diversity, Palestine, Fakuos

Context and objectives: The unmonitored use of herbal medicinal remedies by patients with cancer presents a significant challenge to oncology healthcare professionals. We describe an increasingly popular herbal "wonder drug," Ephedra foeminea (Alanda in Arabic), whose use has spread from the Palestinian patient population throughout the Middle East. We conducted a multicentered and multidisciplinary collaborative research effort in order to understand the potential benefits and harms of this popular herbal remedy. Methods: We conducted an in-depth search of the medical literature, both traditional and modern, for any mention of the clinical use of Alanda for the treatment of cancer. We then tested the remedy, first for toxic ephedra alkaloid components and then for anticancer effects, as well as effects on the cytotoxic activity of chemotherapy agents (cisplatin and carboplatin) on breast cancer cell cultures. Results: We found no mention in the literature, both conventional and traditional, on the use of Alanda for the treatment of cancer. Laboratory testing did not find any toxic components (i.e., ephedra alkaloids) in the preparation. However, in vitro exposure to Alanda led to a reduced cytotoxic effect of chemotherapy on breast cancer cell cultures. Conclusions: The use of an integrative ethnobotanical, laboratory and clinical research-based approach can be extremely helpful when providing nonjudgmental and evidence-based guidance to patients with cancer, especially on the use of traditional herbal medicine. The effectiveness and safety of these products need to be examined by integrative physicians who are dually trained in both complementary medicine and supportive cancer care.

Background Although there have been recent advances in the development of animal-attached ‘proximity’ tags to remotely record the interactions of multiple individuals, the efficacy of these devices depends on the instrumentation of sufficient animals that subsequently have spatial interactions. Among densely colonial mammals such as fur seals, this remains logistically difficult, and interactions between animals during foraging have not previously been recorded. Results We collected data on conspecific interactions during diving at sea using still image and video cameras deployed on 23 Antarctic fur seals. Animals carried cameras for a total of 152 days, collecting a total of 38,098 images and 369 movies (total time 7.35 h). Other fur seals were detected in 74 % of deployments, with a maximum of five seals detected in a single image (n = 122 images, 28 videos). No predators other than conspecifics were detected. Detection was primarily limited by light conditions, since conspecifics were usually further from each other than the 1-m range illuminated by camera flash under low light levels. Other seals were recorded at a range of depths (average 27 ± 14.3 m, max 66 m). Linear mixed models suggested a relationship between conspecific observations per dive and the number of krill images recorded per dive. In terms of bouts of dives, other seals were recorded in five single dives (of 330) and 28 bouts of dives <2 min apart (of 187). Using light conditions as a proxy for detectability, other seals were more likely to be observed at the bottom of dives than during descent or ascent. Seals were also more likely to be closer to each other and oriented either perpendicular or opposing each other at the bottom of dives, and in the same or opposite direction to each other during ascent. Conclusions These results are contrary to animal-attached camera observations of penguin foraging, suggesting differing group-foraging tactics for these marine predators. Group foraging could have consequences for models linking predator behaviour to prey field densities since this relationship may be affected by the presence of multiple predators at the same patch.

Rhus coriaria L. (Sumac) is an important and a widely used crop in the Mediterranean basin as a food spice, and also in folk medicine due to its health promoting properties. Phytochemicals present in plant foods are -in part- responsible for these consequent health benefits. Nevertheless, detailed information on these bioactive compounds is still scarce. Therefore, the present work was aimed at investigating the phytochemical components of sumac fruit epicarp using HPLC–DAD/QTOF-MS in two different ionization modes. The proposed method showed powerful for the tentative identification of 211 phenolic and other phyto-constituents; most of which, have not been described so far in R. coriaria fruits. More than 180 phytochemicals (tannins, (iso)flavonoids, terpenoids, etc.) are reported herein in sumac fruits for the first time. The obtained results highlight the importance of R. coriaria as a promising source of functional ingredients, and boost its further use in food and nutraceutical industries.