Establishment Of Suspension Culture Line On Momordica Charantia L Cells

The compositions of Momordica Charantia L are saponins which have efficacy on falling blood sugar, regulating immunity, resisting oxidation, regulating internal secretion, falling cholesterin, and resisting tumour and so on. For the objective of producing saponins by large-scale suspending culture Momordica Charantia L cells, this paper has studied on the establishment of suspension culture line. The cells of suspension culture line in the research were separated from callus of Momordica Charantia L, so the first step of the experiment was inducing callus from seedlings. And to obtain the cells of calluses were loose, broken easily and suitable to the suspension culture line, the calluses needed to culture continue and optimize the culture condition. Then separating cells from the calluses that had been optimized and farther cultured.At the experiment of inducing callus, continuing culture and optimizing the culture condition: using different concentrations of hormones to inducing calluses from Momordica Charantia L seedlings, and comparing their growth circumstances. Educing the conclusion was that the effect on inducing calluses by 6-BA or KT with IAA was better than 6-BA or KT with 2.4-D. When the concentration of 6-BA was higher (4mg/L), it was benefit to induce calluses. Adopting single factor experiment to make sure the factors and levels that could better affect the calluses' forms and growth. And according to the orthogonal experiment to ensure the calluses growth faster and the forms were loose and broken easily. Educing the conclusion was that basing on MS culture medium added 103.125mg/L ammonium nitrate, 100mg/L hydrolyzed casein, 30g/L sucrose, 0.5mg/L 6-BA, and 0.2mg/L 2.4-D, could proliferate the callus rapidly.Plant cells suspension culture is that cells or cell congeries are cultured and suspended in liquid culture medium, the cells or cell congeries can be cultured in triangles on shake bed, also can be cultured cosmically in biology reactor. Successful cells suspension culture line should have 3 postulates: first, the cells or cell congeries is dispersed state well; second, uniform all right; third, grow speedily. A nicer suspension culture line can increase a great deal in a short time. Adopting single factor experiment to make sure the factors and levels that are suitable for the cells growth, And according to the orthogonal experiment to find out the best level in which the cells growth and saponins anabolism well. The conclusion: the primary and secondary factors that affected Momordica Charantia L cells growth were B (Inoculum's concentration) >D (Combined hormone concentration) >C (Sucrose concentration)>A(Ammonium nitrate affixation concentration), the factors B,D are extraordinary salience. Culture condition was that basing on LS2.5 culture medium added 206.25mg/L ammonium nitrate, 30g/L sucrose, 1.0mg/L 6-BA, 0.5mg/L IAA. and the inoculum's concentration (dry weight) was 4.5g/L. The primary and secondary factors that affected Momordica Charantia L saponins anabolism were C (Sucrose concentration) >A (Ammonium nitrate affixation concentration) > B (Inoculum's concentration) >D (Combined hormone concentration) , the factors A,C were tarnal salience, factor B was extraordinary salience. Culture condition was that basing on LS2.5 culture medium added 103.125mg/L ammonium nitrate, 50g/L sucrose, 0.5mg/L 6-BA, 0.2mg/L IAA, and the inoculum's concentration was 4.5g/L.