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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.

Hibiscus sabdariffa L. has been used traditionally in many countries of the world as food, especially as a flavouring agent in food industry. H. Sabdariffa is used for treating heart, nerve, liver disease, high blood pressure, arteriosclerosis, sore throat, cough, hypoglycaemia, laxative, diuretic, kidney stone, scurvy... The aims of this study are evaluation of bioactivities of H. Sabdariffa and the production of transformed hairy root of H. Sabdariffa for pharmaceutical production. In this study, Yen and Duh method showed the reducing power of ethanol the root extract and leaf extract are higher than that of stem. The root extract showed the α-glucosidase inhibitory activity with IC50 at 0.2 mg/mL is higher than those of stem and leaf. These results shows that root has higher bioactivites than stem and leaf. In this study, hairy roots of H. Sabdariffa were successfully induced via Agrobacterium rhizogenes ATCC 15834 in the plant cells. The frequency of hairy root and number of hairy root induction from the wounded sites of leaves are the highest (100% and 12.89 roots). The stable introduction of rolB and rolC genes of A. rhizogenes ATCC 15834 into H. Sabdariffa plants was confirmed by PCR analysis. Besides, the absence of virG gene confirmed hairy roots as bacteria-free. Subsequently, these results demonstrated that H. Sabdariffa, particularly the roots, has great potential as pharmacological values and hairy root production can be used as pharmaceutical sources.

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One of the most effective methods for type 2 diabetes treatments is inhibition of enzyme α-glucosidase in the intestines to slow down the release of glucose from carbohydrates in the diet, reduce plasma glucose levels and prevent hyperglycemia after meals. Therefore, seeking α-glucosidase inhibitors used in the treatment of diabetes from plant is the attention of many scientists. Based on the potential of the hairy root culture technology in increasing valuable chemical compounds accumulating, this study aimed to induce hairy roots from six plants of the Malvaceae family including Urena lobata, Abutilon indicum, Hibiscus Sabdariffa, Hibiscus rosa-sinensis, Sida acuta, Sida rhombifolia, and screening which materials has the highest in α-glucosidase inhibitory activity. We have successfully induced hairy roots from six plant species by using the Agrobacterium rhizogenes ATCC 15834 strain. The highest rates of hairy root induction were observed in Hibiscus Sabdariffa and Urena lobata. The stable introduction of rolB and rolC genes to plant genomes was confirmed by PCR. Under liquid-shake culture conditions on MS medium, hairy roots of Hibiscus sabdariffa, Urena lobata and Sida acuta showed better development than other species, and therefore, they are selected for the study of α-glucosidase inhibitory activity. This study proved that Urena lobata was stronger in inhibiting α-glucosidase activity than other studied plants, with the IC50 value of 7.65 μg/ml. The results of this study demonstrated Urena lobata hairy root might be considered as a potential supply of medicinal plants for the treatment of type 2 diabetes.