The growth, adventitious bud formation, bioactive flavonoid production, antioxidant response, and cryptochrome-mediated light signal transduction in Isatis tinctoria L. hairy root cultures exposed to LED lights

Abstract

The root of Isatis tinctoria L. is highly appreciated as a Traditional Chinese herbal medicine for the prevention and adjuvant treatment of respiratory diseases caused by coronaviruses viruses such as SARS and COVID-19. I. tinctoria hairy root cultures (ITHRCs) provide a better alternative to field cultivation for the production of antiviral flavonoids. For the first time, ITHRCs were exposed to different colors of LED lights i.e., red, green, blue, red/green/blue (1/1/1, RGB), and white, in an attempt to promote the root growth and enhance the production of bioactive flavonoids. Results revealed that the biomass productivity (7.15 ± 0.63 g/L) in ITHRCs with an initial inoculum size of 0.2% cultured for 50 days under blue light increased by 1.86-fold relative to that under dark (control), and yields of rutin (320.49 ± 27.56 μg/g DW), quercetin (388.75 ± 9.17 μg/g DW), kaempferol (787.90 ± 83.43 μg/g DW), and isorhamnetin (269.11 ± 20.08 μg/g DW) increased by 4.15-fold, 9.31-fold, 9.09-fold, and 2.88-fold as compared with control, respectively. Interestingly, the emergence of adventitious buds was noticed in ITHRCs under all light treatments. Additionally, the enhanced densities of chloroplasts and root hairs were found in blue-light grown ITHRCs as against control, which might account for the elevated biomass productivity. Moreover, blue light induced oxidative stress in ITHRCs in terms of the overproduction of oxidation products and the enhancement of antioxidant enzyme activity. Furthermore, blue light significantly activated photoreceptor (CRY1) and key regulator of light signaling (HY5), thus leading to the up-regulated expression of MYB4 and structural genes (such as CHS and FLS) responsible for flavonoid biosynthesis. And, the transcriptional activation of CUC1 was likely related to the formation of adventitious buds in ITHRCs. Overall, the simple supplementation of blue LED light makes ITHRCs more attractive as plant factories for obtaining higher productivity of biomass and medicinally important flavonoids.