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目的分析腐胺和过氧化氢调控白桦三萜合成中是否存在交互作用。方法在白桦悬浮细胞的生长末期添加1 mmol/L腐胺(Put)、0.1 mmol/L过氧化氢(H2O2)和40μg/mL真菌诱导子,采用比色法、高效液相色谱法和荧光显微镜分析白桦悬浮细胞中三萜量、多胺量和H2O2荧光强度变化。结果 1 mmol/LPut增加了细胞中H2O2的荧光强度和三萜量。0.1 mmol/LH2O2促进了三萜合成,却抑制了多胺中亚精胺(Spd)、精胺(Spm)的合成,而对Put无影响。Put与H2O2同时处理组,细胞中H2O2的荧光强度和多胺的量均介于Put与H2O2单独处理组之间,而三萜量却显著高于其单独处理组;进一步药理学实验分析发现,真菌诱导子与H2O2清除剂过氧化氢酶(CAT)处理后,细胞中的Put和Spd量比真菌诱导后增加了0.62%和16.05%,而对Spm无显著影响。真菌诱导子与多胺(Pas)合成抑制剂D-精氨酸(D-arg)处理组,细胞中的H2O2荧光强度比真菌诱导子处理组有所降低了;在真菌诱导子、CAT和D-arg同时处理下,细胞中的H2O2荧光强度、PAs量和三萜量均低于真菌诱导子与CAT或D-arg处理,但三萜量仍高于对照组。结论 Put与H2O2在促进白桦三萜合成中存在交互作用。
Objective To analyze the interaction between putrescine and hydrogen peroxide in the regulation of triterpene synthesis in white birch. Methods 1 mmol / L putrescine, 0.1 mmol / L hydrogen peroxide (H2O2) and 40 μg / mL fungal elicitors were added to the suspension of Betula platyphylla at the end of growth. The colorimetric method, high performance liquid chromatography and fluorescent microscope The amount of triterpene, the amount of polyamines and the change of fluorescence intensity of H2O2 in the cell suspension of Betula platyphylla were analyzed. Results 1 mmol / LPut increased the fluorescence intensity of H2O2 and the amount of triterpene in the cells. 0.1 mmol / LH 2 O 2 promoted the synthesis of triterpenoids, but inhibited the synthesis of spermidine (Spd) and spermine (Spm) in polyamines, but had no effect on Put. Put and H2O2 treatment group, the fluorescence intensity of H2O2 and the amount of polyamines were between Put and H2O2 alone treatment group, while the triterpenoid content was significantly higher than the treatment group alone; further pharmacological experimental analysis found that, After fungal elicitor and H2O2 scavenger catalase (CAT) treatment, the amount of Put and Spd in the cells increased by 0.62% and 16.05%, respectively, after fungi induction, but had no significant effect on Spm. The fluorescence intensity of H2O2 in fungi and fungi induced by D-arginine (D-arginine), a fungal elicitor, and a polyamine (Pas) At the same time, the fluorescence intensity, PAs content and triterpene content of H2O2 in cells were lower than that of fungal elicitors and CAT or D-arg treatment, but the content of triterpene was still higher than that of control group. Conclusion There is an interaction between Put and H2O2 in promoting the synthesis of triterpene in white birch.