Study On Tissue Culture Of Medicinal Plant Shan-wu-gui

Shan-wu-gui commonly refers to a type of Stephania plants with tubers.Most Shang-wu-gui plants distribute in two major areas of China,huazhong area and southwest.The tubers are rich source of alkaloids and have a series of applications in traditional Chinese medicine(TCM) or folklore herbs.It is usually used to treat fever,inflammation,sedation and acesodyne.Alkaloids research of Shan-wu-gui plants have advanced rapidly in the past few years.Especially,developed separation and purification technologies and high sensitivity detection instruments have been employed in isolating alkaloids from Shan-wu-gui plants.So far,dozens of alkaloids have been separated from different Shan-wu-gui plants,and their physical activities are clarified constantly. Shan-wu-gui boasts a huge potential in market demand.But,owing to the low efficiency of reproduction in natural conditions,the habitat fragmentation and unduly commercial collections, Shan-wu-gui plants have become rare and endangered in recent years.This study optimizes the tissue culture and rapid-proliferation techniques of Shan-wu-gui,thus laying a basic foundation for commercial producing large-scale seedlings,relaxing the contradiction between the Shan-wu-gui supply and demand.The conclusions are listed as follows:1 Tender stem segments was optimal for explant primary culture.The best sterilizing method for tender stems was disinfected with 70%ethanol for 15 s,then treated with 0.1%HgCl₂ appending one or two drop of Tween-80 for 6 min,the ratio of survival was 73.6%.2 Through researching the effect of different hormone combination on budding rate and shoot growth,we found that the medium of MS + NAA 0.1 mg/L + 6-BA 1.0 mg/L was superior hormone combination,which could be used in the first generative cultivation.The average length of plantlets reached 3.5 cm after 35 days' cultivation.3 Subproliferation needed to be cultivated alternately in the following three media:MS + IBA 0.4mg/L + NAA 0.2 mg/L + GA₃ 0.5～1.0 mg/L,MS + NAA 0.1 mg/L + GA₃ 1.0 mg/L,MS + IBA 0.2mg/L + 6-BA 0.4～0.8mg/L + GA₃ 0.5～1.0mg/L,propagation coefficient was 7 times/50d.4 Roots were induced on the medium of 1/2 MS+IBA 0.5 mg/L,which was added 0.06% active carbon.The rooting ratio was 87.4%.The better basal material was sand:pearlite(1:1) and the ratio of survival was 84%.