Fig. 6

Hairy roots of OVX-BcEPSPS in I. indigotica. (a) Schematic representation of the BcEPSPS overexpression vector. T35S, Bar, P35S, CaMV 35S promoter, flag tag, P35S, hygromycin resistance gene. (b) Induction and culture of I. indigotica hairy roots in different periods. a, A sterilised plant of I. indigotica. b, Agrobacterium tumefaciens C58C1 stains carrying plasmids were applied to infect the sterilized leaf. c, The hairy roots were derived from sterile leaves after 2 weeks. d, The false positive hairy roots without the vector C58C1-OVX-BcEPSPS on the 1/2 MS solid medium were supplemented with a gradual decrease in cefotaxime and hygromycin. e, The positive hairy roots containing the vector C58C1-OVX-BcEPSPS showed great growth on the 1/2 MS solid medium with cefotaxime and hygromycin. f, Liquid cultures of positive hairy roots containing the vector C58C1-OVX-BcEPSPS in an Erlenmeyer flask in 45 days. (c) Molecular identification of transgenic hairy roots. Lane 1, DNA size marker, lane 2, the engineered strain as the positive control, lane 3, wild-type control. Lane 4–11, strains of OVX-BcEPSPS. The primers hpt, rolb and rolc were used to check the hygromycin resistance and plasmid Ri. (d) Growth rates of WT, CK and OVX-BcEPSPS hairy root lines