叶子诺 翟明霞 周文洋 肖婉莹 詹亚光 范桂枝

摘要 [目的]探究硫化氫（H2S）对白桦悬浮细胞中吲哚乙酸（IAA）、脱落酸（ABA）以及多酚和三萜等次生代谢物累积的影响。[方法]采用酶联免疫吸附法、实时荧光定量PCR和比色法等分析IAA和ABA含量及其关键酶基因表达水平、黄酮和三萜等次生代谢物含量。[结果]H2S供体硫氢化钠（NaHS）处理对白桦悬浮细胞中IAA、ABA的累积存在浓度和时间效应，其中1.0 mmol/L H2S供体NaHS处理12 h时ABA含量达最高值，比对照增加了196.46%，而IAA含量在0.1 mmol/L H2S供体NaHS处理3 h时达最高值，比对照增加了5.55%。ABA或H2S处理后白桦悬浮细胞中次生代谢产物积累呈增加趋势，而IAA处理则相反。1.0 mmol/L H2S供体NaHS与不同浓度的IAA和ABA分别处理白桦悬浮细胞24 h后，不同程度地提高了白桦悬浮细胞中多酚、黄酮、三萜含量。其中，1.0 mmol/L ABA与 NaHS共同处理后白桦悬浮细胞中多酚、黄酮、三萜含量均达到最大值，分别比NaHS单独处理增加了17.89%、14.74%和8.87%。[结论]H2S处理促进白桦悬浮细胞中IAA、ABA和次生代谢物的累积，H2S与IAA或ABA共同处理可进一步促进次生代谢产物的累积。

关键词 H2S;白桦;IAA;ABA;次生代谢产物

中图分类号 S792.153 文献标识码 A 文章编号 0517-6611（2021）17-0117-05

doi：10.3969/j.issn.0517-6611.2021.17.032

Abstract [Objective]The purpose of this study was to clarify the effect of hydrogen sulfide （H2S） on the accumulation of indoleacetic acid （IAA）， abscisic acid （ABA） and secondary metabolites in birch （Betula platyphylla Suk.） suspension cultures.[Method]The colorimetric method， real-time quantitative PCR and enzyme-linked immunosorbent assay were used to analyze the contents of flavones， polyphenols， triterpenes， IAA and ABA， and the gene expression level of IAA and ABA-related key enzyme.[Result]The results showed the concentration and time effects of H2S donor sodium sulfide （NaHS） on the accumulation of IAA and ABA in the birch suspension cells. Among the cells， the highest level of ABA was at 1.0 mmol/L H2S donor NaHS after 12 h treatment， with increasing rates of 196.46% compared to the control.And the highest level of IAA was at 0.1 mmol/L H2S donor NaHS after 3 h treatment， with increasing rates of 5.55% compared to the control. The accumulation of secondary metabolites in the birch suspension cells showed an increasing trend after ABA or H2S treatment， while the opposite was observed after IAA treatment. Treatment on birch suspension cell with 1.0 mmol/L H2S donor NaHS and different concentrations of IAA and ABA for 24 h increased the polyphenols， flavones and triterpenes levels， respectively. Polyphenols， flavonoids and triterpenes reached maximum levels after combined treatment with 1.0 mmol/L ABA and NaHS; the levels increased by 17.89%， 14.74% and 8.87%， compared to NaHS treatment， respectively.[Conclusion]H2S treatment induced the synthesis of IAA and ABA and the accumulation of secondary metabolites in the birch suspension cells. Co-treatment with H2S and IAA or ABA respectively， could further promote the accumulation of secondary metabolites.

Key words H2S;Betula platypylla Suk.;IAA;ABA;Secondary metabolites

白桦（Betula platyphylla Suk.）为桦木科（Betulaceae）桦木属（Betulalinn）的落叶乔木，适应性强，在我国分布范围广[1]。白桦富含三萜类、黄酮类、多酚类等次生代谢物，其中白桦酯醇和白桦酯酸等三萜物质是白桦的代表性次生代谢物，具有抗菌、抗病毒、提高机体免疫力、抗肿瘤等多重功能[2-4]。为了安全并可持续地利用白桦次生代谢物资源，笔者前期利用组织培养技术建立了白桦悬浮细胞培养体系，并证实白桦悬浮细胞中含有白桦酯醇、白桦酯酸和齐墩果酸等三萜物质、多酚和黄酮等次生代谢物。……