Tissue Culture Of Isodon Amethystoides A Genuine Regional Drug Of North Anhui Province

Isodon amethystoides(Benth)CyWuet Hsuan,belonging to Isodon genus of Lemiaceae family,is a plant species endowed with various medicinal efficacy like clearing heat,detoxification,anti-virus.It is therefore known as a “natural plant antibiotic”.In Suzhou region of Anhui Province.For folk customs,I.amethystoides has been used for the treatment of various diseases,such as tumors,malignant sores,scabies,pulmonary tuberculosis,lung abscess,nephritis,chronic bronchitis,pharyngitis,hepatitis B and psoriasis.I.amethystoides contains abundant of terpenoids [(C5H8)n],with the tetracyclic diterpene and its derivatives as the major constituents.Currently,the I.amethystoides drugs used are usually the wild resources,making the wild resources decreased day by day.Due to the shortage of seedling supply,large scale artificial cultivation has not been achieved yet.By using the cell totipotency,tissue culture technique may provide sufficient I.amethystoides seedlings to realize the large scale cultivation.In this study,we investigated the effect of different plant growth regulators on aseptic culture and propagation of I.amethystoides via orthogonal design,and screened out suitable medium formula for I.amethystoides seedling production under in vitro conditions.The main results are as follows.1.Explants types influence callus initiation time and callus quality.Compared with other explant types,leaf could generate high quality callus.The optimal medium formula for callus induction was MS + 1.5 mg·L-16-BA + 0.5 mg·L-1NAA+ 30 g·L-1sucrose + 7.0 g·L-1 agar.2.The suitable medium formula for callus differentiation was MS +2.0 mg·L-16-BA+ 0.6mg·L-1IBA +2.0 mg·L-1KT + 30 g·L-1sucrose + 7.0 g·L-1agar.3.With plant height,leaf number and adventitious bud induction rate as indicators,an orthogonal experiment was designed to investigate the plant growth regulators on micropropagation of I.amethystoides seedlings.The optimal medium formula was obtained as MS + 1.0 mg·L-1 6-BA +1.0 mg·L-1 IAA + 32 g·L-1 sucrose +7.0 g·L-1 agar,where the adventitious differentiation rate and induction rate were 1.33 and 0.75 respectively.