Rhodiola Hairy Root Culture System To Establish

To establishment of hairy roots cultures of Rhodiola sachalinensis A.Bor with Agrobacterium rhizogenes strains (A4,R1601, and ATCC15834), the following experiment parameters and procedures were optimized, such as selecting some suitable explants, the growth condition for explants, the period of Agroinfiltration, and the growth condition for hairy roots. The research results showed that hairy roots of Rhodiola sachalinensis A.Bor were induced with A. rhizogenes strains (A4,R1601, and ATCC15834), the strains (A4 and ATCC15834) of which had the better effect. It has also been found that the leaf as explants was superior to these organisms. The bacterium concentration OD600=0.4-0.5 and AS (50μmol/L) to the medium for 25 minutes increased the infiltration frequency. The infection frequency was generally higher when the co-cultivation durations of 2 days and co-cultivation medium at pH 5.0 was applied. The addition of Cb (150㎎/l) and Cef (100㎎/l) to the medium showed the optimum resistant-bacterium efficiency. PCR and PCR-Southern blot revealed the integration of rolC gene from Ri T-DNA into the plant genome. The transformed roots grew rapidly on hormone-free medium and showed typical hairy root phenotype. The optimum growth conditions of hairy roots were identified as follows: 1/2 MS0 liquid medium, light free, temperature 20oC, pH 4.5-4.8.