Studies On The Cell Cultures And The Active Material Of Chaenomeles Speciosa Nakai

Tissue and cell culture were used in this study. Different parts of the Chaenomeles speciosa organization were used as the experimental material. In order to induce vigorous growth, structure, loose callus. We establish cell suspension cultures, and research its regulation of the synthesis of secondary metabolites. Systematic studies were used in the inducing of the Chaenomeles speciosa callus. We research the conditions of callus induction, the method to control the loose callus, and the suspension culture characteristics and growth curve of cells. Spectrophotometry was used to determine the changes of the content of OA. It provide experimental and theoretical basis in the industrial production of OA.In the period of the callus induction, the different sources of explants、different sterilization time ratio、the different light conditions were compared on the declared rate of callus induction, browning and growth status of the situation. The results showed that: in the induction medium MS+2,4-D (1mg mL^-1)+KT (0.1mg mL^-1), young stems and leaves are most likely to induce callus. Use these materials, and compare for the disinfection time, the best disinfection time for leaves:75%alcohol5s and0.1%mercuric chloride5min, the best disinfectant time for stem:75%alcohol5s and0.1%mercuric chloride10min. Weak light conditions are favorable for the induct of Chaenomeles speciosa callus.In the period of the callus subculture, loose and strong growth of callus should be obtained. The results status should be adjusted for the providing of materials of the cell suspension culture. The result showed that the subculture of the best recipe: MS+2,4-D (1mg mL^-1)+KT (0.5mg mL^-1) on the subculture, the most suitable sugar concentration is30g L^-1, The best pH is5.5to6.0, the promotion cell subculture time is around20d. Comparison of the light conditions on callus growth, draw a dynamic growth curve during30days. Callus stage in this culture has emerged as "S"-type growth curve, the light condition was more conducive to cell growth, but the browning was more serious in the latter part.Establish the cell suspension cultures of the Chaenomeles speciosa. In order to study the effects of inoculum size, carbon sources, sucrose concentration, pH change during cultivation and other factors on the impact of cell suspension culture, we also determined the dynamic process of cell growth curves and the content of active substances. The results showed that: the MS basic medium with no agar, formulated into liquid medium, the most suitable inoculum was4g flask^-1, most conducive to cell growth carbon source is maltose, but the highest yield of active substances is for the sucrose, so select sucrose as carbon source. The most suitable carbon source concentration is20g L^-1 the active substance production reached0.5g L^-1. The most suitable pH for cell growth is5.5, when cells of the active substance production also reached a maximum for the0.42g L^-1. After5d change the fresh medium in suspension culture can effectively accelerate the rate of cell growth and accumulation of active substances. Comparison of MS medium with different concentrations of suspension cultures, MS is the best medium concentration. Light is significantly promoted on the growth of suspension cells. Establish the growth curve, determinate of cultured cell growth, pH changes and the accumulation of active substances. For the determination of the cycle for30d, during the0⁹d for the lag phase, during the9¹5d for the rapid growth phase, cell growth gradually slowed down since. The pH decreased in the first training cycle, then increased, then decreased during suspension. In order to obtain the content of OA, we use spectrophotometry. And at the27d when the cell content of the active substance and its output has reached the maximum.