硫化氢对白桦悬浮细胞中多胺和次生代谢物积累的影响

王紫晴; 李飞洁; 郭玉婷; 于昭慧; 马钰斌; 周文洋; 范桂枝

doi:10.3969/j.issn.1000-2006.201610011

王紫晴,李飞洁,郭玉婷,于昭慧,马钰斌,周文洋,范桂枝

南京林业大学学报（自然科学版） ›› 2017, Vol. 41 ›› Issue (06) : 26-32.

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南京林业大学学报（自然科学版） ›› 2017, Vol. 41 ›› Issue (06) : 26-32. DOI: 10.3969/j.issn.1000-2006.201610011

研究论文

硫化氢对白桦悬浮细胞中多胺和次生代谢物积累的影响

王紫晴,李飞洁,郭玉婷,于昭慧,马钰斌,周文洋,范桂枝^*

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The effect of H₂S on the accumulation of polyamines and secondary metabolites in birch suspension cells

WANG Ziqing, LI Feijie, GUO Yuting, YU Zhaohui, MA Yubin, ZHOU Wenyang, FAN Guizhi^*

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摘要

【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(NaHS)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】 NaHS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 NaHS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,NaHS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,NaHS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 NaHS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NaHS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NaHS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。

Abstract

【Objective】The purpose of this study was to clarify the effect of hydrogen sulfide(H₂S)on the accumulation of polyamines and secondary metabolites in birch suspension cultures. 【Methods】Different concentrations of H₂S donor sodium hydrosulfide(NaHS)were added to a birch suspension culture. High performance liquid chromatography and colorimetric assays were used to analyze the levels of different forms of polyamines, enzyme activity of arginine decarboxylase(ADC)and polyamine oxidase(PAO), and the levels of flavonoids, polyphenols and triterpenoids. 【Result】 The total polyamine content of birch suspension cultures increased initially and then decreased after NaHS treatment. Among the cells, the highest level of putrescine and spermidine was at 1 mmol/L NaHS after 24 h treatment, with increasing rates of 30.51% and 12.37% compared to the control, respectively. With the exception of perchloric acid(PCA)-insoluble spermidine conjugate, the content of free-, PCA-soluble and PCA-insoluble putrescines and conjugates increased after NaHS treatment. Similarly, NaHS treatment enhanced the activity of ADC and PAO, with increasing ranges of 15.91%-26.33% and 137.5%-250.12%, respectively. Treatment on birch suspension cell with 1 mmol/L NaHS and different concentrations of putrescine, spermidine, and spermine increased the polyphenol, flavonoid and triterpenoid levels, respectively. Polyphenol, flavonoid and triterpenoid reached maximum levels after combined treatment with 5 mmol/L putrescine and NaHS; the levels increased by 26.69%, 13.22% and 4.55%, compared to NaHS treatment, respectively. 【Conclusion】 These results indicate that H₂S treatment increased the levels of polyamines and secondary metabolites, and the combined treatment with H₂S and polyamines further enhanced the accumulation of secondary metabolites in birch suspension cultures.

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王紫晴,李飞洁,郭玉婷,于昭慧,马钰斌,周文洋,范桂枝. 硫化氢对白桦悬浮细胞中多胺和次生代谢物积累的影响[J]. 南京林业大学学报（自然科学版）. 2017, 41(06): 26-32 https://doi.org/10.3969/j.issn.1000-2006.201610011

WANG Ziqing, LI Feijie, GUO Yuting, YU Zhaohui, MA Yubin, ZHOU Wenyang, FAN Guizhi. The effect of H₂S on the accumulation of polyamines and secondary metabolites in birch suspension cells[J]. JOURNAL OF NANJING FORESTRY UNIVERSITY. 2017, 41(06): 26-32 https://doi.org/10.3969/j.issn.1000-2006.201610011

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