Skullcap, Tissue Culture And Hairy Root Induction

Scutellaria baicalensis Georgi is the dry root of one kind of Baicalin Labiatae plants. It has been used for cleaning away heat, moistening aridity, purging fire, detoxifying toxicosis, and is also anti-abortion agent. Because Scutellaria baicalensis Georgi has diverse biological activities, and has highly developed value, it is showing a shortage in the market situation. Because of the increasing scarcity of wild resources, cultivatied Scutellaria baicalensis Georgi appeared. But cultivatied Scutellaria baicalensis Georgi grows slowly, it still can not satisfy the increasing market needs. The application of methods such as tissue culture and hairy roots culture to produce secondary metabolites is feasible and necessary measures.This subject from the biological point of view, applies different induction conditions and culture conditions, selects the best inducing parts, explants, and the induction period. The optimum is the 0.8~1.1cm stem with node, which is growing about 60 days. Simultaneously select the optimal induction conditions: MS+6-BA0.5 mg/L +NAA0.5 mg/L, the illumination condition is 12h/d and so on. On this culture medium, the induction rate of callus of Scutellaria baicalensis Georgi is the highest, it is fresh yellow-green, loose, big, faster and vigorous growth. However, MS+6-BA1.0 mg/L +NAA0.5 mg/L is benefit of bud clusters, they grow vigorously. In this experiment, we also found bud clusters can take root on the culture medium without hormone or only adding NAA.Adding precursors compounds, elicitors, examine that different types and different concentrations of precursor compounds, elicitors impact on PAL callus of Scutellaria baicalensis Georgi. Adding 1mmol/ L NaAc precursor compound, 0.001g/L yeast elicitor is benefit to improve PAL activity. AgNO3 elicitor can also improve PAL activity. Different precursor compounds, elicitors also have varying degrees of impact on its peroxidase isozymes. In this experiment, we found the color of these band had some difference, and the bands of adding NaAc were more than the other two. This subject using Agrobacterium rhizogenes A4 induced hairy roots of Scutellaria baicalensis Georgi. Materials used in the experiment is direct sterilized Scutellaria baicalensis Georgi explants, and callus of Scutellaria baicalensis Georgi, etc. Induced hairy roots of Scutellaria baicalensis Georgi preliminarily, and compared different methods and inducing conditions. The experiment shows if using the direct sterilized explants, OD600=0.6, pre-culture 2d, co-culture 3d, presenting root time is about 17d, more branches, ageotropism hormone from themselves. If using callus to induce hairy root, OD600=0.4, culture 2d, presenting root time is late and about 30d, the induction rate is low. Result shows using this two experimental materials to induce hairy root of Scutellaria baicalensis Georgi still need further discussion.In this experiment, we isolated cell clusters of protoplasts in callus of Scutellaria baicalensis Georgi and compared different experimental conditions. Result shows this is the optimum condition:1%Cellulase R-10,1%Pectolyase,and Macerozyme0.25%, enzymatic hydrolyze 3h, then 1000rpm centrifugate 5min. After adding sucrose 800rpm, centrifugate 5min. Protoplasts can be used as another good experimental material to induce hairy roots of Scutellaria baicalensis Georgi. Although we haven't isolated uniparted protoplast, but conditions can offer some foundations for following experiments.