Improving hairy root induction of Urena lobata L. by Agrobacterium rhizogenes ATCC 15834 by some factors

Abstract

Introduction:Our previous study showed that Urena lobata L. hairy root is a potential pharmaceutical source for type 2 diabetes treatment. In order to improve the transformation efficacy and the quality of hairy roots, this study examined the effects of several factors including age, parts of plants, infection time and culture medium in inducing hairy roots in Urena lobata L. Methods: In this study, we investigated four factors to improve the hairy root induction in Urena lobata L. These factors include: age of plant (15-day-old in vitro plants, 45-day-old in vitro plants and after two subculture generations plants), different parts of plant (roots, stems, and leaves), infection time (10, 20 and 30 minutes), and culture medium (Murashige and Skoog (MS), Gamborg B5 medium (GB5) and Woody plant medium (WPM)). All experiments were repeated three times, with uninfected leaf explants of 15-day-old in vitro as the negative control. The transformation frequency and the fresh biomass of hairy roots were recorded at four weeks after infection. Results: The results showed that the optimized procedure which used 15-day-old in vitro plants, the leafy part, the infection time of 10 minutes and culture in the WPM medium was better than the original procedure. The optimized procedure achieved a transformation frequency of 100%. In addition, the fresh biomass of hairy roots formed on an explant in the optimized procedure was 3.2 times higher than the ones induced by the original procedure. Conclusion: The results showed that the optimized procedure was more effective than the original procedure in inducing Urena lobata hairy roots.

Anti-inflammatory compounds produced in hairy roots culture of Sphaeralcea angustifolia

“…These results are inferior to those reported for other species of the Malvaceae family transformed with A. rhizogenes ATCC 15834 strains. In 15-day-old seedlings shoot explants of Althaea o cinalis, a transformation frequency of 83 ± 5.2% was reported after 5 days of co-cultivation, and in 15-days-old seedlings leaf explants of Urena lobate L. the transformation frequency was 97.33% and 86.33% for stems explants (Phuong et al, 2018;Tavassoli and Afshar, 2018). These relatively low frequencies could be due to the age of S. angustifolia explants, co-cultivation time, and the competence for transforming the plant species.…”

“…There are no reports in the literature about the genetic transformation of S. angustifolia, so studies of species of the Malvaceae family transformed by A. rhizogenes ATCC 15834 strains were used as a starting point. Among such species, we found Gossypium hirsutum and Gossypium barbadense (Triplett et al, 2008); Abutilon indicum L. (Indian mallow; Sajjalaguddam and Paladugu, 2016); Althaea o cinalis L. (marshmallow; Tavassoli and Sa pour Afshar, 2018); and Urena lobata L (Phuong et al, 2018). Based on the above, we decided to establish a transformation protocol for S. angustifolia using A. rhizogenes strains and select hairy root lines with a good growth rate and a high production of scopoletin, tomentin, and sphaeralcic acid, as an industrially competitive biotechnological system.…”

Investigation of factors in improving Agrobacterium-mediated gene transfer in Ruellia tuberosa L. and evaluation of α-glucosidase inhibitory activity in established hairy roots

“…The explants were then immersed in the bacterial suspension (OD600nm = 0.6) for 10 min without adding AS. After the time of infection, the infected explants were incubated in the dark on free-hormone MS medium for 4 d. They were subsequently washed rigorously before being transferred to the MS medium containing 200 mg.L -1 cefotaxime for 30 d (Phuong et al, 2018).…”