Senecio glaucus subsp. coronopifolius (Maire) C. Alexanderis wild annual herb distributed in the Egyptian deserts. Total phenolic and flavonoid content of plant root were determined using both HPLC and colorimetric analysis. Syringic acid and hesperidin (1378.802 and 6638.247 mg / 100 gm. dried plant root powder, respectively) were of the highest concentration compounds resulted from HPLC analysis of total phenolic and flavonoid content. The colorimetric estimation of total phenolic and flavonoid content resulted in concentration of (98.23 ± 0.28 mg/gm. expressed as Gallic acid equivalent (GAE) and35.9± 0.17 mg/gm.expressed as quercetin equivalent (QE), respectively). GC-MS analysis of un-saponifiable matters and fatty acid methyl esters of the plant leaves indicated that octacosane (11.85%) and linolenic acid methyl ester (31.07%) (poly- unsaturated fatty acid) were the major identified compounds, respectively. The DNA of the plant was analyzed using twelve random decamer primers. A total of 52 random amplified polymorphic DNA (RAPD) markers were identified. Root extracts (ethyl acetate, acetone and methyl alcohol) were subjected to determine the antimicrobial behavior and also their cytotoxic activity, by using (3- (4, 5-dimethylthiazolyl-2)-2, 5-diphenyltetrazolium bromide) (MTT) assay against colon carcinoma cell lines (HCT-116). Among the fore mentioned extracts, root ethyl acetate extract gave appreciable antibacterial and antifungal behavior and also had promising cytotoxic activity with IC₅₀ = 7.39 ±1.2  µg/ml. Root methyl alcohol extract showed antioxidant activity with IC₅₀ = 79.57 ± 0.74 µg/ml, using 2, 2-diphenyl-1-picrylhydrazyl (DPPH) assay.