| Fritillaria cirrhosa D.Don is a famous and precious traditional Chinese medicinal plant belonging to the Liliaceae family. In Chinese herbal practices, in which the bulb is generally eaten, it is highly effective for the treatment of relieving cough, removing phlegm, reducing fever and moistening lungs. It is especially effective in the treatment of the elderly and children, particularly in cases of difficult recovery after a lengthy treatment. As a result, the demand of supply in the Chinese drug market is always high. But because of restricted growth requirements and unlimited digging, its nature resources are close to drying up. Considerable efforts in the tissue culture of Fritillaria cirrhosa have been successful. Herbs have character of huge shape after being polyploided, medicinal composition and growing mass are increasing and satisfy the produce of medicinal materials preferably. Combining tissue culture with colchicines treatment induce autopolyploid success in many medicinal plants. Now the tissue culture technique of Fritillaria cirrhosa is perfect, but its polyploid research is vacant. We induce the polyploid of Fritillaria cirrhosa combining tissue culture with colchicines treatment primarily in the article.Calli were induced by culturing germfree bulb in the MS medium supplemented with 2.0mg/L 6-BA and 1 .Omg/L NAA. The diploid wound callus was treated by adding colchicines into the medium and by immersing into colchicines solution for different concentrations and periods. Colchicines concentrations were four levels of 100,500,1000 and 2000 mg/L, and treatment periods were of 1, 5 and10 days for the former and 24,48 and 72h for the latter. The results showed that both the two methods can induce polyploid cells, but the former was better than the latter in the induction rate and operating. The induced effects on Fritillaria cirrhosa callus treated by colchicines with different concentrations and different time were varying, and were better in 500-1000 mg/L with 5-10 days. 1000 mg/L with 5 days was the best and its doubling efficiency was up to 70%. The callus treated was culturing in the MS medium supplemented with 2mg/L NAA to get the root. Microscopic examination of callus and shoot tips showed that the chromosome number of tetraploid was 2n=4x=48, while that of diploid was 2n=2x=24.In this paper, we also study the change of peroxidase isozymes of the callus treated by adding colchicines into the medium and the regenerated tetraploid plants. The results showed that callus and the plants had different regulated modes on peroxidase. Compared with the control, the band number and activity of peroxidase isozymes of callus treated with colchicines were changed obviously, and two new enzyme bands appeared, and some bands disappeared, while the regenerated tetraploid plants only showed the activity raising. So the regenerated tetraploid plant and its diploid were different in inheritance.By the upper research, the best induction method of Fritillaria cirrhosa's polyploid and appropriate concentration and periods of colchicines were obtained, the tetraploid plants were got and identified. The acquaintance of polyploid was deeper via the morphological and cytological examination and peroxidase isozymes.
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