Study On Establishment Of Cell Suspension Culture System And Alkaloid Biosynthesis Regulation Of Lycoris Radiate

Lycoris radiate is a perennial herbs of Amaryllidaceae. The bulbs of Lycoris radiate have many kinds of alkaloid, such as Lycorine, Galanthamine and Lycoramine. Galanmine is a reversible cholinesterase inhibitor. Galanthamine can be treated in Alzheimer’s, Myasthenia gravis, Paralytic ileus, Polio sequelae. Lycorine have treatment of anti-inflammatory, and anti-SARS virus. Lycoris radiate has great prospect because of Galanthamine. But Lycoris radiate has low propagation coefficient in nature, people used Lycoris radiate in traditional methods always from filed. We have lower and lower Lycoris radiate in filed. There is much research on rapid propagation of Lycoris radiate, but the rate of rapid propagation is low too. The cost of getting Lycoris radiate alkaloid is too high depend on rapid propagation. In this article, research on tissue culture and Lycoris radiate alkaloid were summaried. we discovered a new method which depended on cell suspension culture to get Lycoris radiate alkaloid. There was no article to reported on cell suspension culture of Lycoris radiate. The following results were obtained.1Induce callus tissues of Lycoris radiate. Some factors for callus induction had been studied such as various homrone regimes, different parts of bulb and different months. The explants selection was the most significant factor of callus induction. Bulb slices of Lycoris radiate can induce callus tissues, but the inner bulb slices are the best explants for callus inducing, and have better quality. The bestl mediums for the callus induction was MS+2,4-D1mg-L"1+6-BA1mg-L-1and the rate reached61.13%.In growth cycle of Lycoris radiate,the bulb of Lycoris radiate in September and October have higher callus induction rate, and lower in march and can differentiate easier.2Callus subculture of Lycoris radiate. Callus differentiates easily on callus subculture. We can inhibit differentiation to change hormone. The optimal mediums for the callus subculture was MS+2,4-D0.5mg-L-1+KT0.5mg-L-1,and the callus differentiation is difficult. The subculture cycle was24-27days in dark. Light can reduce subculture cycle.The subculture cycle was18-21days in light. Lycoris radiate cell growth curves was "S"-shaped,and can divide into lag phase, logarithmic growth phase and stable phase.3Effects of alkaloid content on callus for light culturing. Light can increase the content of Lycorine, Galanthamine and Lycoramine. In the growth cycle, the content of alkaloid reduce in the beginning. The alkaloid content increased in the late stage of the growth phase. In the end of the cycle, the alkaloid content was approximately equal to the beginning of callus cycle.4Establishment of effective regeneration system of Lycoris radiate.The optimal mediums for budding of callus was MS+6-BA2mg·L-1+2,-D1mg·L-1,and had more quantity of small bulbs. The optimal mediums for rooting of small bulbs was MS+NAA3mg·L-1,and the rate can reach100%.The plants propagation coefficient can crease depend on callus, and the cycle can reduce. The plants grewth well after acclimatizing and transplanting.5The establishment of cell suspension of Lycoris radiate. Some factors for cell suspension had been studied such as inoculums amount, hormone, inositol and condition culture. We discovered the best method of cell suspension which inoculuns amount was15percentage, the medium was MS+2,4-D0.5mg·L-1+KT0.1mg·L-1inositol amount was300mg·L-1.The cell suspension had great quality, the cell suspension had great dispersion, the cell viability was high, cell division was fast. The medium from growth phase can reduce the cell suspension cycle, and the cycle was7days. The cell suspension curves was "S"-shaped, and the cycle can divide into lag phase, logarithmic growth phase and stable phase.The alkaloid content of cell suspension was high. The Galanthamine content was4.18times of the bulb of Lycoris radiate in field.6Effects of alkaloid content of cell suspension for adding precursors. Some research have confirmed, which alkaloid of Lycoris radiate has the same precursors, phenylalanine and tyrosine. Adding phenylalanine had on significant of alkaloid content. Adding tyrosine had significant compared the control group, tyramine too. Adding phenylalanine and tyrosine together could increase alkaloid content, but had no significant compared to add tyrosine only.