Induction Of Fritillaria Ussuriensis Maxim Tetraploid And Study On The Physiological And Biochemical

Fritillaria ussuriensis Maxim as one of the most genuine medicinal materials, this plant are used as clearing away heat and eliminating sputum, relieving cough, anti-inflammation and relieving pain, which has always great concerns on the traditional chinese medicine. As the factors of unique growth environment, longer growth cycle, the yield is lower and the price is so high. However, because of the mankind’s excessively exploiting, the wild resources of this plant has turned into an endangered population. The method of tissue culture could solve these problems.Taking stem of Fritillaria ussuriensis as explants, induce the hormone-self-support callus. Used the hormone-self-support callus as material to induce polyploidy of Fritillaria ussuriensis, to identifie and obtain the tetraploid of Fritillaria ussuriensis. Cultivated the diploid and polyploid of Fritillaria ussuriensis for plant. Sometime,to observe and investigate some physiology and biochemistry of Fritillaria ussuriensis with the different time. The result showed:1. The induction of Fritillaria ussuriensis callus:the explants is the stems of Fritillaria ussuriensis; the optimal culture conditions:the temperature is19℃,dark culture; the optimal medium:MS+2,4-D2.5mg/L2. Hormone-self-support Fritillaria ussuriensis culture system was constructed: Firstly, halved the original subculture hormone concentration2-3times that in MS me dium without hormones, then Fritillaria ussuriensis cultures without hormones were obtained, dark conditions can maintain good growth, The light and shade taking turn i nduce bulbs generating.3. The induction of Fritillaria ussuriensis polyploidy:used the colchicines immersion method, the optimal conditions:the colchicines are500mg/L, time is24h.4. The cellular level and morphological observation: Fritillaria ussuriensis diploid chromosome examination were2n=22, tetraploid were4n=44; Blowhole morphologic observation showed that tetraploid was the giant. And it can obtain the result with plow cytometry.5. The change of some physiology and biochemistry of Fritillaria ussuriensis:The total soluble sugar content showed decrease-increase-decrease-increase trend; the total starch showed to raise steadily; The activity of CAT showed that the trend was decrease-increase and increases gradually; The activity of POD was higher and showed decrease-increase; The activity of PAL was higher in plant, and participated in the accumulation of alkaloid. Over all, the index of tetraploid all higher than diploid.6. Endogenous hormones:GA broke dormancy times in Fritillaria ussuriensis culture, promoted the small bulbs, improved the contention of amylase, and provided energy; IAA participated in formatting of leaf and root; ABA was accumulated in the stage of bulbs for breaking dormancy times.ZR participated in bulbes formation and could promote the formation of root and leaf with IAA. Four types of endogenous hormones will be combined in the growth and development of Fritillaria ussuriensis maxim.7. The alkaloids:The culture of Fritillaria ussutiensis alkaloid congtent is as the same as crude drug or higher than that of crude drug.The alkaloids was smoothed growth in each period.tetraploid of fritillaria ussuriensis alkaloid content is higher than diploid...