Determination And Histochemical Localization Of Alkaloids Content In Wild And Tissue Cultured Fritillaria Cirrhosa D. Don

Beimu ( Fritillaria ) are bulbs of many species of plants from Fritillaria L. and are traditional Chinese drugs. They commonly used as antitussive and expectorant. It is reported that there are 61 species, 50 varieties, and 5 metatypics from Fritillaria. ChuanBeimu ( F. crrihosa ) is a important traditional Chinese drug in Sichuan .province. However, contents of alkaloids are lower in bulbus Fritillaria L. and monomer separating and purification are more difficult. Furthermore, composing of alkaloids from different bulbus Fritillaria L. is also complicated. In this situation, quality evaluation can hardly achieve satisfactory progress. In order to further study, develop and utilize resources of Chinese crude drug in homeland and supply scientific basis for modernization of Chinese herbal medicine, we take studies as follows.The authors research the best supplies of nutrient medium for growing, differentiation and derivation. We mensurate the total alkaloid content of tissue cultured Fritillaria cirrhosa D. Don in different parts and under different growing states, and apply slicing-up technology to histochemically localize the alkaloids, preliminarily build an approach to observe alkaloids distribution with paraffin slices.The experiment result suggests that: in the culture of Fritillaria cirrhosa D. Don,the medium supplemented with NAA(1mg/L) and 6-BA(0.5mg/L) testified the best grouping to make the most callus. NAA with low level is good for growing of callus but on the contrary high-level NAA inhibit it. The tissue culture of Fritillaria cirrhosa D. Don prefer low-level sucrose to high-level. Ribavirin(20mg/L) is good for bulb dense growth someway.The total content of alkaloids of 7 samples (from high to low) was in this order: the tissue cultured regenerative bulb, callus with adventitious roots, browning callus with adventitious roots, callus, browning callus, bulbs of wild Fritillaria cirrhosa D. Don and street drugs of Fritillaria cirrhosa D. Don. The tissue cultured self-born bulb enjoys the most alkaloids and the highest rate of dry/fresh(%), followed by callus with adventitious roots.It is more effective to localize alkaloids with paraffin slices and histochemical approach; alkaloids of bulbs mainly distribute in parenchyma cells while alkaloids of callus mainly around the cell wall; there are relations between alkaloids distribution and starch grains in wild Fritillaria cirrhosa D. Don.In conclusion, it is possible to produce alkaloid by tissue culture, but still need further research.