Utilization of a modern genome mapping technique toinvestigate the impact of artemisia herba-alba microbiome variability on the levels of plant secondary metabolites / Maram Ahmed Adel Eldemerdash ; Supervised Gehad Genidy Mohamed , Mahmoud Saleh Mahmoud , Mohamed Atia Omar

Thesis (M.Sc.) - Cairo University - Faculty of Science - Department of Biotechnology

Artemisia herba-alba is one of the most commonly known effective medicinal plant species that is used worldwide to treat several human and animal ailments. The potential therapeutic effects of A. herba-alba is attributed to the presence of a broad spectrum of bioactive metabolites. The present study aimed to determine the effect of soil microflora on plant secondary metabolites levels and how this could affect its use in medical purposes. Three samples of soil and plant materials were collected from two distinct locations: Mersa-Matrouh (M1, M2, M3) and Saint Catherine (S1, S2, S3). The soil samples were subjected to physicochemical analysis beside total microbiome DNA extraction. Then genomic DNA was extracted from the aerial parts of the plant materials and amplified via SCoT-PCR to determine genetic relatedness of the same plant species from different locations. Plant secondary metabolites were extracted by methanolic extraction and purified using high performance liquid viii chromatography (HPLC). The components were then assessed for biochemical assays including antioxidant, cyclooxygenase inhibitor (COX), lipoxygenase inhibitor (LOX), acetylcholinesterase, and amyloid beta peptide activity assays. Finally, the extracted DNA samples from soil were subjected to 16S rRNA gene amplicon sequencing, and analyzed via Illumina MiSeq. The soil chemical analysis revealed a slightly alkaline pH for all samples and variation in the level of cations between the two locations. SCoT-PCR associations indicated genetic variance within the species and close association between (S2 & S3) and (M1 & M2). HPLC major peaks showed a significance variation in the metabolite profiles of the plants from different locations. Biochemical assays showed for S1-S3; high COX1 and COX2 activity when compared to the control and to M1-M3 samples. Also, high antioxidant activity for M1-M3 when compared to the control and the other samples. Next Generation Sequencing analysis revealed that the most abundant Phyla were Spirochaetes, the most prominent order was Rhodospirillales, Family was XVII, class was Alphaproteobacteria, Genus was Thermaerobacter for all the tested samples

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