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[摘要]
目的 研究外源腐胺对白桦Betula platyphylla悬浮细胞中蔗糖代谢与三萜积累间的关系。方法 在白桦悬浮细胞的生长末期添加0.1、1、5 mmol/L腐胺,采用比色法分析白桦悬浮细胞内蔗糖,果糖和葡萄糖量、蔗糖磷酸合成酶和蔗糖合成酶活性以及三萜量。为进一步明确腐胺与蔗糖对白桦三萜积累的关系,分析蔗糖(5、10、20、40 g/L)和1 mol/L腐胺+5~40 g/L蔗糖处理对白桦三萜的影响。结果 腐胺处理提高了白桦悬浮细胞内蔗糖量,而降低了培养液中的蔗糖量。其中,5 mmol/L腐胺处理6 h时增加幅度最大,比对照增加了169.41%。与蔗糖相比,腐胺对细胞内外果糖和葡萄糖量的影响程度较小,除了处理6 h细胞内的果糖和葡萄糖增加外,其余处理时间细胞内的果糖和葡萄糖量基本呈降低趋势。除5 mmol/L腐胺降低了蔗糖合成酶活性外,腐胺处理增加了蔗糖磷酸合成酶和蔗糖合成酶的活性。5~40 g/L蔗糖处理12 h后,悬浮细胞中的三萜量呈增加趋势,其中20 g/L蔗糖处理时增加幅度最大,比对照增加了52.16%。1 mmol/L腐胺分别与不同浓度蔗糖共同处理后,除腐胺与5 g/L蔗糖共同处理三萜量比单独腐胺增加27.37%外,其他处理均降低了三萜的积累。结论 腐胺处理诱导了白桦悬浮细胞中蔗糖的合成和三萜的积累,同时推测腐胺诱导的蔗糖在一定程度上介导了三萜的积累。
[Key word]
[Abstract]
Objective To analyze the effect of exogenous putrescine on triterpenoid accumulation and sucrose metabolism. Methods Putrescines (0.1, 1, and 5 mmol/L) were added to the eight-day-old suspension cell culture, and the content changes of sucrose, fructose, glucose, and triterpenoid, the activity of sucrose phosphate synthase (SPS) and sucrose synthase (SS) were analyzed by colorimetry. Results Putrescine increased the content of sucrose in cells but decreased the conent in culture medium. Among them, the content of sucrose was most enhanced after putrescine induction for 6 h; The increasing rate was 169.41%. However, the contents of fructose and glucose were only increased in 6 h under the putrescine treatment, the other treatments decreased the contents of fructose and glucose. In addition to 5 mmol/L putrescine treatment, putrescine enhanced the activities of SPS and SS. The content of triterpenoid in suspension cells was increased in 12 h after 5-40 g/L sucrose induction, and the content of triterpenoid was most induced by 20 g/L sucrose, the increasing rate was 52.16%. In addition to increasing 27.37% on triterpenoid content in cells treated by 1 mmol/L putrescine and 5 g/L sucrose, the treatment of 1 mmol/L putrescine and other sucrose concentration decreases the accumulation of triterpenoid. Conclusion Putrescine could increase the sucrose content and triterpenoid accumulation in suspension cell culture, and we preliminarily inferred that the induction of sucrose by putrescine could participate in the accumulation of triterpenoid in birch cells.
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[基金项目]
中央高校基本科研业务专项资金项目(DL13EA08-03);国家自然科学基金资助项目(31100445,31070531);黑龙江省博士后科研启动基金(415288)