诱导剂对海巴戟莨菪亭累积的影响

Effect of inducers on scopoletin accumulation in Morinda citrifolia

【目的】研究诱导剂对海巴戟悬浮细胞系及离体叶片中莨菪亭含量的影响,为海巴戟悬浮细胞系大规模培养生产莨菪亭及揭示莨菪亭累积机理奠定基础。【方法】以连续培养10代以上的海巴戟悬浮细胞系及离体叶片为材料,在海巴戟悬浮细胞系中分别添加不同质量浓度的苯丙氨酸、咖啡酸、肉桂酸、对香豆酸、阿魏酸等5种诱导剂,培养不同时间后检测莨菪亭含量,筛选出诱导剂培养的最佳质量浓度和培养时间,并分析海巴戟悬浮细胞系在最佳培养条件下的生长情况。【结果】在海巴戟悬浮细胞系中,苯丙氨酸和咖啡酸均能诱导莨菪亭累积且促进细胞生长,其中50 mg/L苯丙氨酸培养48 h时莨菪亭含量最高,细胞干质量浓度为7.4 g/L;800 mg/L咖啡酸培养6 h时莨菪亭含量最高,达到56.5μg/g,细胞干质量浓度为8.0 g/L;而肉桂酸、对香豆酸、阿魏酸在不同质量浓度及培养时间下均未在悬浮细胞系中检测到莨菪亭,并且容易导致悬浮细胞系褐化。用不同质量浓度诱导剂培养海巴戟离体叶片,苯丙氨酸、咖啡酸、肉桂酸、对香豆酸和阿魏酸诱导海巴戟离体叶片累积莨菪亭的最佳培养质量浓度分别为50,800,500,1 000和800 mg/L,且其分别在培养8,24,36,4和8 h时莨菪亭含量达到较高,依次为1 018.3,1 725.0,1 013.7,848.9和785.3μg/g。【结论】在大规模海巴戟悬浮细胞系生产莨菪亭时,可优先选用800 mg/L咖啡酸作为诱导剂,能在短时间内获得最大量的莨菪亭。

【Objective】This study investigated the effects of inducers on scopoletin contents in Morinda citrifolia suspension cell lines and detached leaves to lay foundation for large scale culture of M.citrifolia suspension cell lines to produce scopoletin and reveal the mechanism of scopoletin accumulation.【Method】Suspension cell lines and detached leaves of M.citrifolia cultured continuously for more than 10 generations were selected and inducers including phenylalanine,caffeic acid,cinnamic acid,p-coumaric acid and ferulic acid were added to suspension cell lines.After different culture time periods,contents of scopoletin were detected,optimal concentration and culture time were determined and the growth of M.citrifolia suspension cell lines was analyzed.【Result】Both phenylalanine and caffeic acid could induce the accumulation of scopoletin and promote cell growth of M.citrifolia suspension cell lines,and 50 mg/L phenylalanine induced the highest scopoletin content at 48 h with cell dry mass concentration of 7.4 g/L,while 800 mg/L caffeic acid induced the highest scopoletin content of 56.5μg/g at 6 h with cell dry mass concentration of 8.0 g/L.Cinnamic acid,p-coumaric acid and ferulic acid could not induce scopoletins in suspension cell lines at different concentrations or culture time periods,and they were prone to lead to browning of suspension cell lines.When culturing M.citrifolia detached leaves,50 mg/L phenylalanine,800 mg/L caffeic acid,500 mg/L cinnamic acid,1000 mg/L p-coumaric acid and 800 mg/L ferulic acid were the optimal concentrations to induce scopoletin accumulation of scopoletin and highest scopoletin contents of 1018.3,1725.0,1013.7,848.9 and 785.3μg/g were produced at 8,24,36,4 and 8 h,respectively.【Conclusion】Caffeic acid at 800 mg/L was preferred as the inducer for producing scopoletin in large scales through M.citrifolia suspension cell lines,which could obtain the most amount of scopoletin in short times.