Genetic Engineering Dept, CINVESTAV-IPN, Unidad Irapuato, P.O. Box 629, C. P. 36500, Irapuato, Guanajuato, México.
Plant Cell Rep. 1996 Jan;15(5):360-6. doi: 10.1007/BF00232372.
The biosynthesis of the sesquiterpenic phytoalexin capsidiol was investigated using in vitro root cultures of chili pepper (Capsicum annuum) elicited with cellulase. Optimal concentrations of cellulase and sucrose for capsidiol production were established. A simple spectrophotometric procedure to quantify capsidiol was improved. Monoclonal antibodies against a tobacco sesquiterpene cyclase were used to detect a similar protein in pepper root extracts. We found that capsidiol was secreted to the medium and the maximal production was achieved at 24 h after elicitation. In contrast, the maximal amount of the elicitor inducible sesquiterpene cyclase was found between 6 and 8 h. Addition of small amounts of polyvinylpyrrolidone was necessary for sesquiterpene cyclase enzyme activity assays.
采用纤维素酶诱导的辣椒(Capsicum annuum)离体根培养体系研究了倍半萜植物抗毒素capsidiol 的生物合成。确定了用于 capsidiol 生产的纤维素酶和蔗糖的最佳浓度。改进了一种简单的分光光度法来定量 capsidiol。使用针对烟草倍半萜合酶的单克隆抗体来检测辣椒根提取物中的类似蛋白。我们发现 capsidiol 被分泌到培养基中,在诱导后 24 小时达到最大产量。相比之下,在 6 至 8 小时之间发现了最大量的诱导性倍半萜合酶。在进行倍半萜合酶酶活性测定时,需要添加少量聚乙烯吡咯烷酮。