Study On The Physiological Regulation Of Secondary Metabolites Of Curcuma Longa L

Curcuma longa L. is treated as a traditional Chinese medicine. Curcumin is themain active ingredient, which has effects of antioxidant, lowing blood fat, anti-tumor,anti-inflammatory and has been treated as a safe and effective medicine with manypharmaceutical activities. This research established turmeric in vitro culture system.And discuss the effect of light mass, elicitor, precursor and NaCl stress on the growthcharacteristics and the physiological regulation of secondary metabolites in turmericin vitro culture system. Results are listed as follows:1. The most suitable culture medium for the propagation and growth of theCurcuma longa’s tissue culture seeding is: MS+6-BA2.0mg·L-1+NAA0.5mg·L-1+TDZ0.05mg·L-1with sucrose concentration of35g·L-1. The robust tissueculture seeding with high growth modulus and good growth condition can beachieved under this condition.2. The leaf of the Curcuma longa’s cannot induce the callus. The base is easier toinduce the callus than stem. The most suitable culture medium for the callus inducingis: MS+2,4-D1.0mg·L-1+6-BA3.0mg·L-1. The most suitable culture mediums forthe subculture micropropagation of Curcuma longa’s callus is: MS+2,4-D1.0mg·L-1＋6-BA1.0mg·L-1＋NAA0.2mg·L-1and MS+2,4-D1.0mg·L-1＋6-BA4.0mg·L-1.The high appreciation rate of Curcuma longa’s callus is got by three time subculturemicropropagations with loosen and disperse callus which can be used for suspensioncultivation’s materials.3. Green light is the most conducive to Curcuma longa’s tissue culture seeding.Under the condition of red light, blue light, red light: blue light=3:1and red light:blue light=7:1, the more chloroplast pigments are synthesized. Yellow light canincrease the content of the soluble sugar to some extent. Except blue light, otherwavelength of light can all inhibit protein synthesis in varying degrees. When redlight: blue light=7:3, the tissue culture seeding grows better with high appreciationrate, robust plant, highest activities of4CL and C4H and more p-coumric acids andferulic acids are synthesized. Blue light can promote the synthesis ofdemethoxycurcumin and curcumin. Red light: blue light=3:1can result in morebisdemethoxycurcumin and demethoxycurcumin. Red: blue=1:1has some promotion in synthesizing ferulic acids and cinnamic acids.4. After adding salicylic acid and sodium nitroprusside, the appreciation rate, theplant height and the diameter of the base stem are decreased sharply that shows thetwo kinds of inducer has some inhibition on Curcuma longa’s tissue culture seeding’sgrowth. The salicylic acid in some concentration can induce more chloroplastpigments, bisdemethoxycurcumin, caffeic acid, ferulic acid and cinnamic acid, whilerestrain the synthesis of demethoxycurcumin, curcumin and coumaric acid to someextent. The content of protein and soluble sugar are increased when the concentrationof the salicylic acid is0.5mg·L-1. The activities of4CL and C4H are increased underthe concentration of the salicylic acid of0.1~0.5mg·L-1. The concentration of sodiumnitroprusside (0.01~1.0mmol·L-1) can inhibit the synthesis of protein, on the otherhand, increase the synthesis of soluble of sugar. When the concentration of thesodium nitroprusside is0.1mmol·L-1, the four organic acids’ content is increased. Thecontent of curcumins are increased when the sodium nitroprusside is1.00mmol·L-1.5. The content of bisdemethoxycurcumin is raising and the content ofdemethoxycurcumin is decreasing while the concentration of caffeic acid is from0.1mmol·L-1to1.00mmol·L-1. When the concentration is0.1mmol·L-1.,it is mostsuitable for the synthesization of curcumin. The highest activities of4CL and C4H arefound when the concentration of phenylalanine is0.5mmol·L-1. We also find that thecontent of bisdemethoxycurcumin reaches at its maximum with the least content ofdemethoxycurcumin and curcumin at the concentration of phenylalanine (2.0mmol·L-1). While the concentration of cinnamic acid is from0.01~0.05mmol·L-1, theactivities of4CL, C4H and the content of bisdemethoxycurcumin,demethoxycurcumin and curcumin are increased gradually with the maximum at the0.2mmol·L-1. After adding ferulic acid, the content of chlorophyll a and chlorophyll bis raising with the most content of soluble sugar、protein and the activities of4CL,C4H synthesized. When the concentration of cinnamic acid is0.01~0.05mmol·L-1,we can get more curcumin. Adding p-coumric acid can get more chlorophyll with themore soluble sugar and protein at the concentration of0.05mmol·L-1and get the mostcurcumin at the concentration of0.07mmol·L-1.6. The content of chlorophyⅡand carotenoid decrease gradually under NaCl stress. The concentration of the NaCl (25～50mmol·L-1) can promote the growth ofCurcuma longa. With the increase of the concentration of NaCl, the content of thesoluble sugar, protein, and the activity of4CL, C4H are decreased after rising at first.All index reach to maximum when the concentration of NaCl is75mmol·L-1,. Theactivities of the PAL,4CL and C4H are higher than control sample when theconcentration is50mmol·L-1. NaCl can increase the content of the four organic acidto some extent but inhibit the synthesis of curcumins.7. The most suitable culture medium for cell suspension culture of Curcuma longais: MS+2,4-D0.5mg·L-1+TDZ2.0mg·L-1with fructose (30g·L-1). Loosen anddisperse cells of Curcuma longa with larger cell biomass with good growth condition,high content of demethoxycurcumin and bisdemethoxycurcumin can be achievedunder this condition. The optimal cycle of the cell suspension culture of Curcumalonga is24days.