Shu-fang Wang1, Yi-ming Sun, Wei Lei, Rui Yang, Min Sun. 1. Key Laboratory of Eco-environments in Three Gorges Reservoir Region Ministry of Education, School of Life Sciences, Southwest University, Chongqing 400715, China.
Abstract
OBJECTIVE: To establish a culturing system of hairy roots of Scutellaria viscidula, and study the hairy roots growth and biosynthesis of flavonoid in the culturing system. METHOD: Hairy roots of S. viscidula were obtained from infected stem explants after infection with the disarmed Agrobacterium tumefaciens strain C58C1, elite strains were screened and growth curves were determined. The transformation of Ri T-DNA was examined through PCR and baicalin content was examined through HPLC. RESULT: Hairy roots appeared in vitro 8 days after inoculation of S. viscidula with disarmed A. tumefaciens strain C58C1. After 24 days the frequency of stems explants was up to 81%. Transformation was confirmed by the amplification of rolC genes from the hairy roots of S. viscidula. The results demonstrated that rolC genes could be expressed in hairy roots of S. viscidula. Under the 36 days suspension culture of S. viscidula hairy roots in 1/2MS medium, dry weight of hairy root increased 17.42 times, the content and baicalin increased 21.60 and 25.56 times. CONCLUSION: The establishment of the culturing system of hairy root of S. viscidula provided a foundation for further industrial production of active drug component.
OBJECTIVE: To establish a culturing system of hairy roots of Scutellaria viscidula, and study the hairy roots growth and biosynthesis of flavonoid in the culturing system. METHOD: Hairy roots of S. viscidula were obtained from infected stem explants after infection with the disarmed Agrobacterium tumefaciens strain C58C1, elite strains were screened and growth curves were determined. The transformation of Ri T-DNA was examined through PCR and baicalin content was examined through HPLC. RESULT: Hairy roots appeared in vitro 8 days after inoculation of S. viscidula with disarmed A. tumefaciens strain C58C1. After 24 days the frequency of stems explants was up to 81%. Transformation was confirmed by the amplification of rolC genes from the hairy roots of S. viscidula. The results demonstrated that rolC genes could be expressed in hairy roots of S. viscidula. Under the 36 days suspension culture of S. viscidula hairy roots in 1/2MS medium, dry weight of hairy root increased 17.42 times, the content and baicalin increased 21.60 and 25.56 times. CONCLUSION: The establishment of the culturing system of hairy root of S. viscidula provided a foundation for further industrial production of active drug component.