| Tripterygium wilfordii Hook f.is the Euonymus Branch vine plants,TheTripterygium as traditional Chinese herbal medicines have more than700yearshistorical ustilize in China.It has a wide range of pharmacological effects,clinicallyusually used for treatmenting to rheumatoid arthritis,lupus,nephritis and so on.Modernmedical researchs show that Tripterygium also has significant anti-inflammatory,anti-tumor,anti-organ rejection,the role of immunomodulatory and anti-fertility.Atpresent,resources of the traditional medicinal plants are still more than the main toexcavation wild resources,unreasonable and destructive use of plant resources,has ledto medicinal resources dwindling.With the demand of medicinal plants resourcesincreasing,plant tissue culture technology to produce medicinal secondary metabolitesof great significance,and also provides a new way to achieve sustainable use oftraditional Chinese medicine resources.Tripterygium tissue vaccine material,the callusinduction,the establishment of suspension cell culture system,the impact ofxenobiotics on biological characteristics of suspended cells and secondarymetabolites.The main results and contens are given as follows：1．The Tripterygium tissul seedling was used to reseach the induction rate ofdifferentt explants and callus Triptolide contents.The results found that the order ofexplants callus induction on MS medium:leaves﹥stems﹥roots;Leaves explantinduction on different components of the culture medium:MS﹥N6﹥B5﹥White﹥1/2MS;Meanwhile,the studies show that tripterygium wilfordii leavescallus induction the best range of pH is5.8to6.2.the qualities of different explantscallus distribution rule: leaf callus﹥root callus﹥stem callus.2．Studied the growth of callus cells and content of Triptolide under differentnutritional conditions after suspension culture of leaf callus of Tripterygium.The resulted indicated that Suspension cells proliferated the fastest in the White medium,while grew the slowest on MS medium.The triptolide content of suspension cells inWhite and MS medium had reached the max both in the first6days,which were7.14mg/g (DW) and4.77mg/g.The maximum yield of triptolide content in N6medium was6.34mg/g appeared in the ninth day.The max content of tirptolide wasbest on the medium of White with28.13ug/ml,and the follow was N6with21.27ug/ml,and MS with16.27ug/ml.Meanwhile the pH value of mediums had fluctuationsphenomenon with the growth of cells.3．Salicylic acid (SA) and lanthanum chloride (Lacl3) have a different role inpromoting of Tripterygium suspended cells PAL activity; Differences in the twoelicitor treatment in suspension cells within the mass fraction of triptolide.Twoelicitors concentration treatments in cells under a peak mass fraction of the totalvariance of the distribution law:0.1mg/LSA95.99μg/g﹥40mg/LLacl386.34μg/g﹥1.0mg/LSA71.02μg/g﹥60mg/LLacl365.47μg/g﹥CK63.12μg/g﹥20mg/LLacl358.23μg/g﹥10.0mg/LSA54.86μg/g.Salicylic acid (SA) andlanthanum chloride (Lacl3) have the ability of promoting to suspension cells to theextracellular release of secondary metabolites in appropriate concentrations,couldpromote the suspension of a pigment cell extracellular accumulation.Results for thetwo elicitors accumulated effect of Tripterygium cells outer a hormone peakproduction： SA0.1mg/L5.53mg/L﹥Lacl320mg/L3.11mg/L﹥Lacl340mg/L2.68mg/L﹥Lacl360mg/L1.84mg/L﹥CK1.56mg/L﹥SA1.0mg/L1.48mg/L﹥SA10.0mg/L1.34mg/L.4．Different times add the endophytic fungi elitors to process cell,the peak massfraction of the overall distribution of Variation:NS-5mycelium⑧155.18μg/g>NS-5mycelium④140.62μg/g> NS-17culture fluid④101.12μg/g> NS-5culturefluid④93.41μg/g> NS-17mycelium④75.52μg/g> NS-17mycelium⑧70.72μg/g>CK63.12μg/g> NS-5medium⑧53.1μg/g> NS-17medium⑧39.4μg/g.Differenttimes of the elicitor on the extracellular peak concentration of A prime effect of thelaw as follows:NS-5mycelium elicitor (K8)> NS-17mycelium elicitor (K6)> NS-17medium elicitor (K5)> NS-5culture fluid elicitor (K4)> NS-5culturefluid elicitor (K7)> NS-17mycelium elicitor (K2). |
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