Ajuga Bracteosa Transgenic Regenerants Display Better Pharmacological Potential

Abstract Ajuga bracteosa Wall. ex Benth is an endangered medicinal herb used against different ailments in folklore medicines. Here, we aimed to create a new insight to the fundamental mechanisms of genetic transformation in the ethnomedicinal usage of this plant. We transformed the plant with rol genes of Agrobacterium rhizogenes and raised the regenerants from the hairy roots. The transgenic regenerants were screened for in vitro antioxidant activities, a range of in vivo assays, and linked the activities with elemental analysis, polyphenol content and different phytochemicals found through HPLC. Among 18 polyphenolic standards, kaempferol was found most abundant in all transgenic lines (up to 101.26 ± 6 µg/mg). Furthermore, among all tested plant extracts, transgenic line 3 (ABRL3) showed maximum phenolics (13.39 ± 2µg GAE/mg) and flavonoids content (4.75 ± 0.16 µg QE/mg). ABRL3 also demonstrated potent total antioxidant capacity (8.16 ± 1 µg AAE/mg), total reducing power, (6.60 ± 1.17 µg AAE/mg), DPPH activity (IC50 = 59.5 ± 0.8µg/mL), hydroxyl ion scavenging (IC50 = 122.5 ± 0.90 µg/mL), and iron chelating power (IC50 = 154.8 ± 2 µg/mL) among all plants. Transformed plant extracts also produced significant analgesic, anti-inflammatory, anticoagulant, and antidepressant properties in in vivo mice model as compared to control untransformed plant material. Additionally, no abnormal behavior or lethality was observed in any animal tested. In conclusion, transgenic regenerants of A. bracteosa pose better pharmacological properties under the effect of rol genes as compared to wild type plants.