从山豆根组培根中提取、分离独脚金属杂草发芽刺激物质的研究

Isolation of a germination stimulant for Striga hermonthica (Del) Benth from root culture of Menispermum dauricum DC

试验研究了从山豆根(Menispermum dauricum DC.)组培根中提取、分离独脚金属杂草[Striga hermonthica(Del)Benth]发芽刺激物质,其提纯、分离过程是首先将发芽剌激物质吸附在XAD-4树脂表面后采用甲醇脱洗,通过乙酸乙脂:水分配提取,活性物质在Sephadex LH20开口型人工填充柱上进行柱层析,活性组分合并后进一步采用商业提供的C18 Sep-Pak(10g)柱进行柱层析,之后采用分取和分析高效液相色谱提纯、分离,每一步提纯过程中均采用Striga种子发芽实验鉴定活性物质的存在。高效液相色谱分析表明有3种活性物质,其中主要活性物质与Strigol有十分相似的色谱特性,最后经质谱鉴定为Strigol或Strigol类似物质,并首次报道Strigol是植物的代谢产物。

In this paper the isolation and purification of Striga hermonthica germination stimulant from Menispermum dauricum DC.root culture filtrate are reported.Purification and isolation were undertaken by absorption of the stimulants onto XAD-4,solvent partitioning into ethyl acetate,and further separation on Sephadex LH20,C18 Scp-Pak Cartidge,preparative and analytical HPLC columns.Each step involved in the isolation procedure was followed by a bioassay using conditioned Striga hermonthica seeds.Three stimulants were detected in the filtrate.On HPLC the major stimulant showed similar chromatographic behavior and co-eluted with authentic strigol.Furthermore,its UV and Mass spectra were similar with those reported for strigol and strigol-like molecules.In this study,albeit no novel Striga germination stimulant was isolated from M.dauricum root culture,the investigation represents the first report on isolation of strigol or strigoHike molecule from tissue culture.Moreover,it provides unequivocal evidence that strigol and strigoHike molecules are of plant origin.