Research On Suspension Culture And Plant Regeneration Of Psammosilene Tunicoides Hairy Roots

Psammosilene tunicoides is an endangered medicine plant in the southwest of China. Compared to other cultures, it has many merits such as high biomass, strong accumulation of active compounds, and stable production, using the method of liquid culture of hairy root from the Agrobacterium rhizogenes-infected explants.In this study, Agrobacterium rhizogenes (ACCC10060) was employed to induce hairy root from the young leaves of Psammosilene tunicoides as explants. Then a liquid culture system for hairy root was established through primary screening of several factors including liquid medium, N sources and P sources, After the liquid culture system of hairy root has been established, improving the inductivity and growth of Psammosilene tunicoides hairy root callus by optimizing plant growth regulators.The main results are as follows:1. In this experiment, two parameters including biomass and total saponins were used as indicators, which employed to evaluate the effects of N sources and P sources on the growth of P. tunicoides hairy roots during liquid culture. When the ratio of NH4+to NO3-was1:2in the culture media, the biomass and total saponins contents both were achieved at their peak points, likewise6.25mmol/L PO43-act in a similar manner in these two respects, the content of total saponins in hairy roots reached to0.84%(DW) and the biomass accumulated to3.358±0.244mg (DW).2. The absorption and usage of NO3-and PO43-by P. tunicoides hairy roots were consistent with the growth cycle of hairy roots, their concentrations in media almost declined to the lowest level at the25th day of culture and then maintained stable status.3. Investigate the efforts of different concentration of hormonal combinations on dedifferentiation and proliferation of hairy root. The results showed that2,4-D1.0mg/L+6-BA1.0mg/L+NAA2.0mg/L was proven to be the best for the hairy root dedifferentiation and the best proliferation medium of the hairy root callus was MS+2,4-D0.5mg-L-1+6-BA1.0mg/L+NAA0.5mg/L+KT0.2mg/L.4. Detection showed that hairy root callus contains opines that retained the Ri plasmid The obtained hairy root callus exhibited some significant properties of fast growth, secondary metabolic active, its relative growth rate was0.0802and content of total saponins reached to0.837%.5. Add different hormonal combinations to the medium, inducing buds from callus of hairy root. The result showed that the most suitable hormonal combination is6-BA2.0mg/L+NAA0.2mg/L.Through the study showed that hairy roots and hairy root callus of medicinal composition accumulation speed are far more than the original plant, with people’s increasing demand for medicinal plants and medicinal resources increasingly scarce, and by hairy root culture technology and cell culture technology instead of traditional Chinese medicine planting has important significance.