Sichuan Rhodiola Tissue Culture And Its Resistance Gene Detection Of The Situation

Rhodiola rosea,a precious Tibetan folk, usually grow in the alpine plateau areas. Recent pharmacological studies showed that the medicinal Rhodiola rosea exhibits strong activities of anti-anoxia, anti-fatigue, anti-toxic, anti-radiation, anti-tumour, anti-aging, and active-oxygen scavenging. In addition, its roots were extensively used to prevent senility and fatigue, as well as improve human immunity. However, the weak propagation capability of the plant seed resulted in wild source sparse. Meanwhile, directly introducing the plants to low altitude area is unproductive, because it is vulnerable to plant diseases and insect pests. Therefore, plant tissue culture and frequent reproduction are urgently needed to protect the resource and the genetic diversity of Rhodiola rosea.Through the technique of plant tissue culture, we obtained the regenerative plant of Rhodiola rosea. Above all, we studied the effects of different nutrition condition on callus inducing, callus differentiation, rooting and influences of different time when explant were sampled and different explant on callus inducing. In addition, we identified the special expression of the resistant genes in several periods of plant tissue culture. The results were showing as following aspects:1. As for the determination of explant, October is the optimal time for sampling the Rhodiola rosea. At that time, callus inducement ratio of explants (stem and leaf) reached 78%. Stem could be induced callus after 5 days, while leaf needs 10 days.2. The plant hormone 2, 4-D and NAA was generally used to induce stem callus, but 2,4-D shows callus inducement earlier than NAA in the earlier stage(one week after inoculated), and the callus appear white pellet and delicate structure. In later stage(two weeks after inoculated), the growth potentiality of callus, appearing light yellow and green, with NAA in the medium was higher than that with 2, 4-D in the medium.3. The optimum concentration of plant hormone, promoting the callus inducement and growth of stem and leaf, were 6-BA 2mg/L, NAA 0.2mg/L, 2, 4-D 2mg/L, KT 0.5mg/L.4. The best recombination of the concentration of plant hormone and sucrose is 6-BA 2mg/L, NAA 0.2mg/L, sucrose 40g/L. High concentration sucrose promote callus differentiates.5. Through identifying the special expression of the resistant genes in several periods of plant tissue culture, 11 special genes were investigated, the result indicated that complicated reactions present in the different processes of plant tissue culture. Further study is needed to explain the mechanism of gene regulation in the process.