| Isatis indigotica Fort.a medicine which is widely used for curing kinds of illness. At the base of father research ,the object of tissue culture study is to exploit the technique systems of rapid propagation and regeneration.Meanwhile,combined with chromosorme engineering, the method and technology on selectively breeding Isatis indigotica Fort.Germplasm has been done for obtaining the optimal breeding mether.In the research, using the asepsis Isatis indigotica Fort.seed and tine of caudex as explant. Keep the seed bringing up by the MS medium with different concentration of phytohormone and cell mitosin to induced callus,callus multiplication,callus polarization,induced buds,buds multiplication and induced the best growing root; At the 25℃temperature,Isatis indigotica Fort.were doubled using 0.05%,0.1%,0.2%,0.5% Colchicines treatment with germinating seeds and tine of caudex in different treatment time at the 6h,12h,24h,48h. Isatis indigotica Fort.doubled plants were studied on aspects of cytology and morphology at their tillering stage.The antisepsis result showed that : It is very important to get the asepsis explant. Because the sterilized effect can affect the growth of caudex and callus. The seeds were sterilized 10 mins by 0.1% Hgcl2 and 30 seconds by 75% alcohol ,rinsed 6～7times by sterilized water, can get 67.5% bourgeon rate and the pollute rate is just 2.5%.The regeneration system of Isatis indigotica Fort.showed that: The optimal culture medium of induced hypocotyl polarization is MS+0.5mglL6-BA, induced rate is 88%; The optimal culture medium of induced cotyledon polarization is MS + 0.5mg/L 6-BA + 0.5mg/L NAA, induced rate is 86%; The optimal culture medium in which induced hypocotyl becomes callus is MS + 1.0mg/L 6-BA + 0.5mg/L 2,4-D, induced rate is 74%; The optimal culture medium in which induced cotyledon becomes callus is MS + 2.0mg/L 6-BA + 0.5mg/L 2,4-D, induced rate is 86%; The optimal culture medium of callus multiplication is MS+1.0mg/L 6-BA+1.0mg/L 2,4-D; The optimal culture medium of callus polarization is MS+1.0 mg/L 6-BA +1.0 mg/L NAA,The polarization rate is 63.3%;The optimal culture medium of buds multiplication is MS+0.5mg/L 6-BA +0.5 mg/L NAA; The optimal culture medium of growing root is MS+0.2mg/LNAA+0.2mg/LIAA,the growing root rate is 88%.The result of cytology and morphology showed that: gemmiferous seeds and tine of caudex treated by 0.2% aqueous solution of colchicines and treatment time in 6h can get the most polyploidy plants. The effect tine of caudex is better than gemmiferous seeds, the double rate of gemmiferous seeds is 36.8%; the double rate of tine of caudex is 45.3%. All doubled plants were mixoploids including digploids and tetraploids . The mutation plant has the characters of the leaf becoming bigger,broader;the colour becoming darker;and the plant becoming lower.The thickness of leaf is 1.59 times as thick as doubled.the length of leaf is 1.44 times as long as doubled,the extent of leaf is 1.76 times as broad as doubled, the leaf index is 69.12% times. The leaves stoma of defending cells and stoma of tetraploids were longer than that of diploids.the stoma of defending cells's length is 1.41times as long as doubled,the stoma of defending cells's extent is 1.40 times bigger than diploids,the stoma's length is 1.59 times longger,the stoma's extent is 1.84 times broader than diploids, In the acreage of unit,the number of stoma is reduced evidently, lower 0.58 times. The number of chromosome increased two times, and the chloroplast increased too, about 1.63 times.
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