| The generation of medicinal plants’ secondary metabolites is closely related to the regulation of environmental factor, and studies reveal that environmental stress could promote the accumulation of secondary metabolites. While Plants is a very complicated system, having various environmental reaction or control system, hence it is difficult to apply plant to secondary metabolites research to some extent. The culture of Medicinal plants cells worked out this problem, playing an critical role in lowering dimension decomposition to some degree. Suspension cultured cells of Sorbus aucuparia was usually applied to research biphenyl secondary metabolites due to its blooming and response delicacy.The program takes Suspension cultured cells of Sorbus aucuparia L. as object, to study the influence of abiotic elicitors heavy metals and biological elicitor yeast extracts on suspension cultured cells of Sorbus aucuparia via measuring cell biomass and change trend of secondary metabolites. The main research contents include:1. Culture Studies on Suspension cultured cells of Sorbus aucuparia2. Research on the influence of abiotic elicitors heavy metals on synthesis and accumulation of cell biomass and secondary metabolites of suspension cultured cells of Sorbus aucuparia3. Research on the influence of biological elicitor yeast extracts on suspension cultured cells of Sorbus aucuparia4. Influence of composite elicitors of heavy metals and yeast extracts on suspension cultured cells of Sorbus aucupariaResearch results:1. The experiment measured the growth cycle of20d and the pH value of cell suspension liquid of suspension cultured cells of Sorbus aucuparia under normal culture conditions, determined the growth cycle of suspension cultured cells of Sorbus aucuparia. Until14d’s culture, the cell fresh weight of suspension cultured cells of Sorbus aucuparia reached the maximum value, during1-5d}s culture, cells are in delay period of growth;during5-14days, cells are in exponential period of growth;during 14-20days, cells are in plateau period of growth. Under normal culture conditions, changes in the pH value of cell suspension are increasing obvious. Via fitting the growth curve of suspension cultured cells of Sorbus aucuparia by using the fresh weight and dry weight of cells, the results show that the Logistic function has maximum RZ value, up to0.969, indicating that the function fit well, and it can describe the growth curve of suspension cultured cells of Sorbus aucuparia. The function is f (x)=5.044/(1+23.34*exp (0.3887*x)). Seeking first and second order derivative of the fuction,we conclude that suspension cultured cells of Sorbus aucuparia attain maximum growth rate on the8th day,and maximum growth acceleration on the5th day. When cells grow until the5th day, the early stage of exponential, cells are fit to elicitor treatment during this period.2. Via utilizing heavy metal to induce suspension cultured cells of Sorbus aucuparia, we have chance to study the changes of cell biomass and secondary metabolites after disposing. The results show that,La, Cu, Pb elicitors have influence of different degree on cell biomass. La elicitor has stimulating effects on cell biomass when the concentration is between lug/L-10mg/L,the stimulating effects show high concentration in the intermediate and low concentration variation trend at both ends. The La concentration of the maximum stimulatory effects on cell biomass is10[g/L (P <0.05).The effects of Cu elicitor on cell biomass is related to induction time,0.05mg/L Cu concentration shows inhibition effect on cell biomass all the time.When induction time <96h, low concentration first show inhibition effect and then promote effect, on the contrary, high concentration first show promote effect and then inhibition effect. Cu concentration of the maximum stimulatory effects on cell biomass is50mg/L (P <0.05).Pb concentration of0.5mg/L,5mg/L,50mg/L show stimulatory effect on cell biomass (P <0.05).There are no secondary metabolites synthesized by suspension cultured cells of Sorbus aucuparia under the inducement by La, Cu, Pb.3. Via utilizing yeast extracts to deal with European suspension cultured cells of Sorbus aucuparia to study the influence of YE on cell biomass, secondary metabolites, pH value of cell suspensions and changes of intracellular soluble protein content. The results show that, under YE inducement,the number of cell biomass showed a decreasing trend,the D-value between theoretical value and the actual value of cell biomass increases with extend of induction time; pH value of cell suspensions shows a decreasing trend;12h after induction, aupuparin can be detected in Europe, intracellular secondary metabolite concentrations changes with induction time varies. The soluble protein content in treated cells is higher than that of the control group,when the induction time <96h, the content is500’0of control group, when the induction time>96h, the content is144%of control group(P<0.05).4. With the treatment of La3+YE composite elicitor,both low and high concentrations of La elicitor show stimulating effect on the increase of total cell biomass, The concentration of0.1g/L,20mg/L of La and lOmg/L of Cu have improvement on inhibition of YE elicitors to cell biomass. the manner and extent that La+YE, Cu+YE, Pb+YE effect Biphenyl compounds is closely related to the concentration.The mode of effect is: Stimulate or inhibit the synthesis of secondary metabolites content; significantly higher the processing cells synthesize a compound maximum value be than that of YE alone induced; advance the peak time that YE induced synthesis of some compound. Cu+YE affects the concentrations of intracellular compounds is related to Cu concentration, Increased or decreased as the concentration of copper increase, during which the inducement time matters. The compound concentration of cells is related to Pb’s concentration, and spell a V shape with a bottom point at YE+0.5mg/L. The compound concentration increases as the Pb concentration increases.Conclusion:1. From the research of suspension cultured cells of Sorbus aucuparia’s growth characteristics, we concluded the function: f(x)=5.044/(1+23.34*exp(-0.3887’`x)), with which the cells’ growth function corresponds. In this case, cells grow fast at a smooth speed in the14th day, and reach the fastest speed in the8th day. During their growth, it is observed the elicitors could be used.2. It is observed that La, Cu and Pb’s concentration have different respectively influence, which have relatively relationship with elicitors’ concentration and the processing time, on the biomass of these cells. In general, cell fresh weight be reflected as Parabolic shape when the La’s concentration increases, while Cu and Pb both as U curve.10[tg/L of La,50mg/L of Cu and0.5mg/Lof Pb could respectively accelerate the increase of biomass to the most. All the three, however, cannot directly cause the formation of Biphenyl compounds, which is made by the suspension cultured cells of Sorbus aucuparia.3. Biphenyl compounds, caused by suspension cultured cells of Sorbus aucuparia, can be producted at a more rapid speed under the influence of YE, which after observation, are2’-OH-aucuparin,3,5-dihydroxy-biphenyl, noraupuparin aucuparin, eriobofuran. Suspension cultured cells of Sorbus aucuparia has specific reaction toward YE, that is to say, stress path should be applied when cells once be stressed.Cell fresh weight is losing profoundly and its rate is increasing when Secondary metabolites being produced. Meanwhile, PH decreases, its soluble protein increases evidently. It is concluded from this case that the PH and Defense protein synthesis may have main relationship with the decreasing of cell fresh weight.4. Although heavy metal can not cause the production of biphenyl compounds, the same effect can be achieved by other ways in dealing with the production of secondary metabolites. During the research of YE and composite induction of three kinds of heavy metal, it is discovered that cell fresh weight increases under the function of induction of La and Cu and cell fresh weight decreases futherly by Pb. It indicates that La and Cu have Antagonistic effect toward the YE stress, Pb, in contrast, has Synergistic effect. The effects by three elements all relate to its concentration. The results arise in96-132h with poor concentration, while48-72h middle concentration and high concentration12-24h. cLanthanum element cannot affect the trend of YE induced secondary metabolite synthesis, but it can increase the production of secondary metabolites. The concentration of1ug/L of lanthanum, can significantly (compared with YE, P <0.05) increased the synthesis of eriobofuran, when the concentration of lanthanum is IOmg/L, there is a marked increase in the volume of the noraupuparin induced by Lanthanum.Cu elements can make the Synthesis peak time of YE induced secondary metabolites the production of secondary metabolites ahead. When the concentration of Cu is0.5mg L, The aucuparin synthesis peak time mentioned from132h to72h. Synthesis of secondary metabolites is related to copper concentration, along with the increase of Cu concentration, secondary metabolite synthesis gradually decreased. For noraupupann,i n the0.5mg/L induced synthesis is6.5times than i n the0.5mglL induced synthesis.The inducing synthesis of YE secondary metabolite is related to the concentration of lead elements.When the concentration is0.05mg/L, O.lmg/L, the trend of Synthesis amount declines, When the concentration is5mg/L,100mg/L, the trend of Synthesis amount raises, When the concentration is5mg/L, Synthesis amount is the lowest. |
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