Establishment of induction and culture system for hairy roots of Psammosilene tunicoides

Abstract

To establish a culture system for Psammosilene tunicoides hairy roots, and provide technological aid for the large-scale production of P. tunicoides material. The young leaves and stem segments of sterile plantlets were infected with ACCC10060 strain, and subsequently a culture system suitable for hairy roots growth was further established. When explants were co-cultured with ACCC10060 (A600 0.8) on B5 media containing 20 mg x L(-1) Acetosyringo (AS) for 48 h, the hairy roots could be successfully induced, and it could achieve a higher induction rate using young leaves as explants than that of stem segments. The transfected hairy roots possessed the ability of kanamycin resistance and growth on hormone-free media, and synthesis of opines. All above results demonstrated that the present hairy roots originated in the infection of P. tunicoides tissues by ACCC10060 strains. After 35 d culture in liquid hormone-free MS (1/2 strength), the biomass of hairy roots increased 14.11 times (fresh weight) and 8. 39 times (dry weight), respectively, and the content of total saponins in hairy roots reached to 0.857% (DW), by contrast, it's only 0.388% and 0.217% in callus and seedlings respectively. Establishment of hairy roots culture of P. tunicoides provided a foundation for industrial production of active components from P. tunicoides culture.