3 research outputs found
Optimization of the Culture Medium Composition to Improve the Production of Hyoscyamine in Elicited Datura stramonium L. Hairy Roots Using the Response Surface Methodology (RSM)
Traditionally, optimization in biological analyses has been carried out by monitoring the influence of one factor at a time; this technique is called one-variable-at-a-time. The disadvantage of this technique is that it does not include any interactive effects among the variables studied and requires a large number of experiments. Therefore, in recent years, the Response Surface Methodology (RSM) has become the most popular optimization method. It is an effective mathematical and statistical technique which has been widely used in optimization studies with minimal experimental trials where interactive factors may be involved. This present study follows on from our previous work, where RSM was used to optimize the B5 medium composition in [NO3−], [Ca2+] and sucrose to attain the best production of hyoscyamine (HS) from the hairy roots (HRs) of Datura stramonium elicited by Jasmonic Acid (JA). The present paper focuses on the use of the RSM in biological studies, such as plant material, to establish a predictive model with the planning of experiments, analysis of the model, diagnostics and adjustment for the accuracy of the model. With the RSM, only 20 experiments were necessary to determine optimal concentrations. The model could be employed to carry out interpolations and predict the response to elicitation. Applying this model, the optimization of the HS level was 212.7% for the elicited HRs of Datura stramonium, cultured in B5-OP medium (optimized), in comparison with elicited HRs cultured in B5 medium (control). The optimal concentrations, under experimental conditions, were determined to be: 79.1 mM [NO3−], 11.4 mM [Ca2+] and 42.9 mg/L of sucrose
The Desirability Optimization Methodology; a Tool to Predict Two Antagonist Responses In Biotechnological Systems: Case of Biomass Growth and Hyoscyamine Content in Elicited Datura starmonium Hairy Roots
International audienceBackground: The use of the desirability function approach combined with the response surface methodology (RSM), also called Desirability Optimization Methodology (DOM), has been successfully applied to solve medical, chemical, and technological questions. It is particularly efficient for the determination of the optimal conditions in natural or industrial processes involving different factors leading to the antagonist responses. Objectives: Surprisingly, DOM has never been applied to the research programs devoted to the study of plant responses to the complex environmental changes, and thus to biotechnological questions. Materials and Methods: In this article, DOM is used to study the response of Datura stramonium hairy roots (HRs), obtained by genetic transformation with Agrobacterium rhizogenes A(4) strain, subjected to the jasmonate treatments. Results: Antagonist effects on the growth and tropane alkaloid biosynthesis arc confirmed. With a limited number of experimental conditions, it is shown that 0.06 mM jasmonic acid (JA) applied for 24 h leads to an optimal compromise. Hyoscyamine levels increase by up to 290% after 24 h and this treatment does not significantly inhibit biomass growth. Conclusions: It is thus demonstrated that the use of DOM can efficiently - with a minimized number of replicates - leads to the optimization of the biotechnological processes