Genetic diversity and volatile oil components variation in Achillea fragrantissima wild accessions and their regenerated genotypes

Background Wild medicinal plants are suffering natural environmental stresses and habitat destruction. The genetic diversity evaluation of wild accessions and their in vitro raised genotypes using molecular markers, as well as the estimation of substances of pharmaceutical value in wild plants and their regenerated genotypes are convenient approaches to test the genetic fidelity of regenerated plants as a source of substances of pharmaceutical value. In this study, the genetic diversity of 12 accessions of the medicinal plant Achillea fragrantissima, representing five sites in the mountains of South Sinai, Egypt, were estimated by the inter simple sequence repeats (ISSR) fingerprinting and their volatile oil components were identified using gas chromatography-mass spectrometry (GC-MS) analysis. The same accessions were regenerated in vitro and the genetic diversity and volatile oil components of propagated genotypes were determined and compared to their wild parents. Results Clustering and principal component analyses indicated that the wild accessions and their regenerated genotypes were genetically differentiated, but the regenerated plants are relatively more diverse compared to their wild parents. However, genetic variation between wild accessions is inherited to their in vitro propagated genotypes indicating genotypic differentiation of the examined accessions. The number of volatile oil compounds in the wild A. fragrantissima accessions was 31 compounds while in the in vitro propagated plants only 24 compounds were detected. Four major compounds are common to both wild and regenerated plants; these are artemisia ketone, alpha-thujone, dodecane, and piperitone. Conclusions Genome profiling and essential oil components analysis showed variations in A. fragrantissima accessions from different populations. Genetic differences between wild and regenerated genotypes were analyzed and validated with the final conclusion that in vitro conditions elicited higher genetic variation that is associated with reduced amount and diversity in the essential oil components.

The natural anti-inflammatory sesquiterpene lactone achillolide A reduces atopic dermatitis in a murine model

Plant-derived substances have been shown to affect potential targets in inflammatory diseases. We have previously purified from the desert plant Achillea fragrantissima, a sesquiterpene lactone named achillolide A, and demonstrated its anti-inflammatory activities in cultured brain macrophages named microglial cells. In the present study, we further investigated achillolide A in alleviating atopic dermatitis, a chronic and recurring inflammatory skin disease. We investigated achillolide A for its in vivo anti-inflammatory activity using the oxazolone model of atopic dermatitis in mice, in which oxazolone induces ear swelling. Our results show that mice treated with achillolide A showed a significant decrease in the oxazolone-induced ear swelling. Since macrophages are inflammatory cells that play a pivotal role in the pathogenesis of atopic dermatitis, the anti-inflammatory effects of achillolide A were also studied in spleen cells. We demonstrated that achillolide A reduced the levels of LPS-induced inflammatory cytokines IL-2, IL-6, TNFα, IFNγ and IL-12 that were secreted from cultured splenocytes. These data suggest that achillolide A should be considered for further research in treating atopic dermatitis.

EFFECT OF BIOFERTILIZATION AND SOME SINAI’S FLORA EXTRACTS ON THE VEGETATIVE GROWTH AND YIELD OF BROAD BEAN

Broad bean plants (Vicia faba L.) were cultivated in two field experiments at the Experimental Station of the Faculty of Environmental Agricultural Sciences, Arish University, North Sinai, Egypt, during 2018/2019 and 2019/2020 seasons under North Sinai condition, to study the effect of soil application of some biofertilizers and foliar spray with some wild plant extracts on broad bean plants. Biofertilizer treatments contained combination of Arbuscular Mycorrhizal Fungi (AMF) + Trichoderma harzianum + Rhizobium leguminosarum. Three wild plant extracts treatments; i.e., Qeysoom Gebeli (Achillea fragrantissima L.), Harmal (Peganum harmala L.) and Mitnaan (Thymelaea hirsute L.) were sprayed on plants. Treatments were arranged in a randomized complete block design with three replicates in split plot system. The results indicated that biofertilizers had significant effected of all studied traits in both seasons. Foliar spraying of Thymelaea hirsute extract had the highest stimulation effects on spores count, root colonization, mycorrhizal status, and weight of non-active nodules, broad bean plant growth characters; i.e. stem length, number of branches per plant, leaf area, and shoot fresh and dry weight as well as both of fresh and dry weight of roots. Both of Thymelaea hirsute and Achillea fragrantissima extracts combined with biofertilizer treatment had significant effects on yield and its components (pod length, number of pods per plant, average pod weight total yield per plant, and weight of seeds per pod) in both seasons. The combination between Thymelaea hirsute and biofertilizer was the superior interaction treatment of this study.

Phytochemical Screening, Antioxidant and Antibacterial Activities of Pollen Extracts from Micromeria fruticosa, Achillea fragrantissima, and Phoenix dactylifera

Pollen is a male flower gametophyte located in the anthers of stamens in angiosperms and a considerable source of compounds with health protective potential. In the present work, phytochemical screening was carried out as well as analysis of the antioxidant and antibacterial properties of pollen extracts from Micromeria fruticosa, Achillea fragrantissima, and Phoenix dactylifera growing wild in Palestine. Phytochemical screening examined the total flavonol, flavone and phenolic content. The DPPH (1,2-Diphenyl-1-Picrylhydrazyl) and FRAP (ferric reducing antioxidant power) methods were used to assess antioxidant propriety, and disc diffusion, minimum inhibitory and bactericidal concentration tests were used to test the pollen extract’s antibacterial activity against multidrug-resistant (MDR) clinical isolates. The highest level of total phenolic was found in the extract of Micromeria fruticosa (56.78 ± 0.49 mg GAE (Gallic Acid Equivalent)/g). The flavone and flavonol content of samples ranged from 2.48 ± 0.05 to 8.03 ± 0.01 mg QE (Quercetin Equivalent)/g. Micromeria fruticosa pollen with IC50 values of 0.047 and 0.039 mg/mL in the DPPH and FRAP assays, respectively, showed the greatest radical scavenging action. In addition, this pollen showed a mild antibacterial action against the microorganisms studied, with MICs varying from 0.625 to 10 mg/mL and inhibition diameters ranging from 13.66 ± 1.5 to 16.33 ± 1.5 mm.

Ethanol Extract of Achillea fragrantissima Enhances Angiogenesis through Stimulation of VEGF Production

Objective: Achillea fragrantissima L. (Asteraceae) is a traditionally used medicinal herb in the rural communities of Jordan. Methods: The present study evaluated the efficacy of the ethanol extract of this species on angiogenesis in both, ex vivo using rat aortic ring assay and in vivo using rat excision wound model. Results: In concentrations of 50 and 100 µg/ml, the ethanol extract showed angiogenic stimulatory effect and significantly increased length of capillary protrusions around aorta rings of about 60% in comparison to those of untreated aorta rings. In MCF-7 cells, the ethanol extract of A. fragrantissima stimulates the production of VEGF in a dose-dependent manner. 1% and 5% of ethanol extract of A. fragrantissima containing vaseline based ointment was applied on rat excision wounds for six days and was found to be effective in wound healing and maturation of the scar. Both preparations resulted in better wound healing when compared to the untreated control group and vaseline-treated group. This effect was comparable to that induced by MEBO, the positive control. Conclusion: The results indicate that A. fragrantissima has a pro-angiogenic effect, which may act through the VEGF signaling pathway.