Mechanism Study Of Elicitation Responses At Different Culture Stages In Taxus Suspension Culture

Today, plant cell culture technology is considered as one of the most promising methods for plant secondary metabolites production. Elicitation can effectively improve the synthesis of secondary metabolites. Given the convenience, economy and effectiveness, elicitation has been widely studied and utilized. In order to optimize time for elicitation and to better improve the elicitation efficiency, we studied the law of susceptivity of Taxus cells in culture cycle at the signal level. From the work below, we have gained some progress:1. Through detecting the variation of p-ERK activities by Western blotting and ROS concentrations by DCFDA during culture cycle, we found p-ERK activities and ROS concentrations varied during culture cycle in"M"type. p-ERK activities'change somewhat earlier than ROS concentrations.2. Through eliciting Taxus cells at different culture stages with 1mM Ce⁴⁺ and detecting the intracellular p-ERK activities and ROS production, we found the cells at different culture stages react differently towards the same elicitor. According to the effect of elicitation, culture cycle can be divided into 3 periods: 1⁸d: Long and strong p-ERK and ROS expressions could be induced after elicitation; 9¹5d: Short and strong p-ERK and ROS expressions could be induced after elicitation; 16²5d: Long and relatively weak p-ERK and ROS could be induced after elicitation. From these characteristics, 9¹5d, especially 9¹1d is the best period for elicitation.Conclusion: The different p-ERK and ROS variational characteristics may be the key reason for different effects induced at different culture stages. Treating 9-11d old Taxus suspension cells with Ce⁴⁺ could induce a fast, prominent and strong cellular signal, and the short-term signal is good for cell viability recovery. This may be the reason that elicition in this period can obtain the best effect.