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目的明确内源多胺是否介导真菌诱导子诱导白桦三萜的合成。方法将40μg/mL真菌诱导子、1 mmol/L腐胺(Put)和2 mmol/L多胺合成抑制剂D-精氨酸(D-Arg)添加到培养8 d的白桦悬浮培养体系中,利用HPLC法和比色法分析多胺量和三萜量。采用药理学和恢复实验分析多胺在真菌诱导子诱导白桦三萜合成中的作用。结果真菌诱导子或Put处理后,白桦悬浮细胞中的多胺量、三萜量和产量均呈增加趋势,其中处理24 h时,三萜量达最大值,分别增加了68.54%和30.34%。真菌诱导子和Put共同处理虽提高了三萜量,但其产量却低于真菌诱导子单独处理。真菌诱导子和D-Arg共同处理后三萜量低于真菌诱导子单独处理,处理24 h时降低程度最高,为40.57%。恢复实验发现,随着恢复时间的延长,真菌诱导子、Put以及真菌诱导子与D-Arg对白桦三萜合成的影响逐渐减弱,恢复到对照水平。结论多胺介导了真菌诱导子促进白桦悬浮细胞中三萜的合成。
Objective To determine whether endogenous polyamines mediated fungal elicitor-induced synthesis of white triterpenoids. Methods 40 μg / mL fungal elicitor, 1 mmol / L putrescine and 2 mmol / L polyamine synthesis inhibitor D-arginine were added to the suspension culture system of Betula platyphylla for 8 days. Polyamines and triterpenes were analyzed by HPLC and colorimetric methods. Pharmacological and recovery experiments were used to analyze the role of polyamines in fungal elicitor - induced synthesis of birch white triterpenoids. Results The amount of polyamine, triterpene and the yield of triterpene in suspension cells increased with the treatment of fungal elicitor or Put, and the maximum value of triterpenes reached 68.54% and 30.34% at 24 h. Co-treatment with fungal elicitor and Put increased triterpene production, but its yield was lower than fungal elicitors alone. The amount of triterpenoids co-treated with fungal elicitor and D-Arg was lower than that of fungal elicitors alone, with the highest reduction at 40 h after treatment of 40.57%. The recovery experiment showed that the fungal elicitor, Put, fungal elicitor and D-Arg on the triterpene synthesis gradually weakened and recovered to the control level with the recovery time prolonged. Conclusions Polyamines mediated fungal elicitors to promote the synthesis of triterpene in suspension cells of Betula platyphylla.