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【目的】分析硫化氢(H2S)对白桦悬浮细胞中多胺含量和次生代谢物积累的影响。【方法】将不同浓度的H2S供体硫氢化钠(Na HS)添加到白桦悬浮培养体系中,采用高效液相色谱法和比色法分析不同形态的多胺含量、多胺合成酶和分解酶活性以及黄酮、多酚和三萜等次生代谢物含量。【结果】Na HS处理后白桦悬浮细胞中总多胺含量呈先增加后降低趋势,其中1 mmol/L Na HS处理24 h时腐胺和亚精胺总含量达最高值,分别比对照增加了30.51%和12.37%;除高氯酸不溶性结合态亚精胺外,Na HS处理后白桦悬浮细胞中游离态、高氯酸可溶性结合态和不溶性结合态腐胺和亚精胺含量均有不同程度的增加;同样,Na HS处理后多胺合成酶(ADC)和多胺分解酶(PAO)活性基本呈增加趋势,增加幅度分别为15.91%~26.33%和137.50%~250.12%;1 mmol/L Na HS与不同浓度的腐胺、亚精胺和精胺分别处理悬浮细胞,不同程度地提高了白桦悬浮细胞中多酚、黄酮和三萜含量。其中,5 mmol/L腐胺与Na HS共同处理后白桦悬浮细胞中多酚、黄酮和三萜含量达最大值,分别比Na HS单独处理增加了26.69%、13.22%和4.55%。【结论】H2S处理提高了白桦悬浮细胞中多胺和次生代谢物含量,H2S与多胺共同处理可进一步促进次生代谢物的累积。
【Objective】 The purpose of this study was to analyze the effects of hydrogen sulfide (H2S) on the accumulation of polyamines and secondary metabolites in suspension cells of Betula platyphylla. 【Method】 Different concentrations of sodium hydrogen sulfite (Na HS) were added into the suspension culture system of Betula platyphylla. The contents of polyamines of different forms, polyamine synthetase and degrading enzyme were analyzed by high performance liquid chromatography and colorimetric method. Activity and secondary metabolites such as flavonoids, polyphenols and triterpenes. 【Result】 The results showed that the total polyamine content in the suspension cells of Naïve Betula platyphylla first increased and then decreased, and the total content of putrescine and spermidine in the 1 mmol / L Na HS treatment reached the highest value, which increased respectively 30.51% and 12.37%, respectively. In addition to perchloric acid-insoluble bound spermidine, the content of free and perchloric acid bound and insoluble bound putrescine and spermidine in the suspension cells of NaHS were different Similarly, the activities of polyamine synthetase (ADC) and polyamine degrading enzyme (PAO) of Na HS increased with the increase of 15.91% ~ 26.33% and 137.50% ~ 250.12%, respectively. HS and different concentrations of putrescine, spermidine and spermine were treated with suspended cells, respectively, to varying degrees, increase the level of polyphenol, flavonoids and triterpene in the suspension cells of Betula platyphylla. Among them, the concentrations of polyphenols, flavonoids and triterpenoids in the suspension cells of Betula platyphylla co-treated with 5 mmol / L putrescine and Na HS were the highest, which were respectively increased by 26.69%, 13.22% and 4.55% than that of Na HS alone. 【Conclusion】 H2S treatment increased the contents of polyamines and secondary metabolites in the suspension cells of Betula platyphylla. Co-treatment with H2S and polyamines further promoted the accumulation of secondary metabolites.