【目的】探索新型诱导剂褐藻酸寡糖诱导大豆生成大豆抗毒素(glyceollins)的最佳条件和累积规律。【方法】采用制备型高效液相色谱的方法对经褐藻酸寡糖溶液诱导的大豆乙醇提取液进行分离纯化,并用超高压液相色谱-串联质谱法(UPLC-MS)对其组分进行确认。在此基础上研究褐藻酸寡糖的浓度、大豆的预浸泡时间、诱导过程中的大豆培养温度及湿度、黑暗中的培养时间对glyceollins积累量的影响,确定褐藻酸寡糖诱导产生glyceollins的最佳条件。【结果】褐藻酸寡糖可以作为外源诱导剂来诱导大豆累积生产glyceollins,当诱导剂褐藻酸寡糖的浓度为4%,大豆在无菌水中浸泡5 h,且经诱导的大豆在温度25℃、湿度60%、黑暗中培养4 d时,大豆中的glyceollins生成量达到最高,为0.525 mg.g-1鲜豆重。【结论】本试验结果为大豆高品质加工和褐藻酸寡糖的进一步开发利用可提供试验研究基础。
【Objective】The objective of this study was to investigate the relationship among alginate oligosaccharides(AOS) elicitation and glyceollins accumulation in soybean.【Method】The preparative high performance liquid chromatography was selected to separate and purify the ethanol extracts of elicited soybeans seeds with AOS,the separation process was monitored by HPLC and UPLC-MS was used to detect their molecular weights.The optimal parameters were determined by investigating the effects of the concentration of AOS,the soaking time,the temperature,moisture and time course.【Result】AOS induced glyceollins synthesis in soybean seeds.The optimal conditions of elicitation were as follows: concentration of AOS 4%,presoaked in the sterile water for 5 h,stored at a controlled temperature of 25℃ and humidity of 60% in the dark for 4 d.Results from the optimal conditions in the study indicated that the maximum concentration of the phytoalexin glyceollins was 0.525 mg·g-1 fresh weight.【Conclusion】 These findings will give some science suggestions for soybean high value processing and the development of AOS.