Ri-mediated transformation of Phytolacca acinosa with GUS gene

Hairy roots were induced from Phytolacca acinosa leaves after co-cultured with Agrobacterium rhizogenes Ar1334 harbouring agropin type pRi1855 and binary vector pIG121HM.The root with many root hairs and branches grew vigorously towards all directions on MS medium without any plant hormone.Leaf discs pre-cultured on MS medium for two days were dipped into bacteria solution delusioned 5 times at exponential phase for 5 min,and then transferred onto fresh MS medium supplement with 500 mg·L~(-1) carbenicillin to eliminat Ar1334,and 20 mg·L~(-1) Kan to selected the transformants.A highest frequency as (82.2%) of root formation was achieved at the optimum transforming conditions.A 560 bp DNA fragment was amplified from the genomic DNA of hairy root by PCR using primes of rol C gene which was one of the four genes related to hairy root induction located on TL-DNA of Ri plasmid.Blue dots were discovered on the hairy root tips after they were dipped into X-gluc solution at 37℃ for one night.These results proved phytolacca acinosa hairy root could be an efficient culture system to express foreign gene and accumulate valuable substance.