独活提取物抑制柑橘病原真菌活性成分的分离与鉴定

Isolation and identification of active compound from Radix Angelicae Biseratae extracts against citrus fungus pathogens

【目的】为了探明独活抑制柑橘病原真菌的活性成分。【方法】采用生长速率法、溶剂萃取、硅胶柱层析、气相色谱-质谱联用(GC-MS)等方法研究独活乙醇提取物对3种柑橘贮藏病害病原真菌的抑菌作用,并分离和鉴定其抑菌活性成分。【结果】独活乙醇提取物中含有高效抑制柑橘炭疽菌的活性物质。GC-MS鉴定B2组分中含有4种香豆素类化合物—异佛手柑内酯、牛防风素、茴芹内酯、异茴芹内酯。异佛手柑内酯、茴芹内酯、异茴芹内酯3种化合物对柑橘炭疽菌的EC50在45 mg.L-1左右,其中茴芹内酯的抑制活性最好,其EC50为41.521 6 mg.L-1。显微观察发现,茴芹内酯处理导致柑橘炭疽菌菌丝体分支增多、畸形,部分隔膜消失等变化。【结论】异佛手柑内酯、茴芹内酯、异茴芹内酯为独活抑制柑橘病原真菌的主要活性物质。

[Objective]The objective of the study is to elucidate the activity compounds from Radix A ngelicae Biseratae against citrus fungus pathogens. [Method]Antifungal virulence of Radix A ngelicae Biseratae ethanol extracts against three kinds of citrus fungus pathogens was measured with growth rate method. Then solvent extraction and silica gel column chromatography was done to separate and purify active ingredients. Fraction B2 was analyzed by GC-MS. [Result]The results showed that ethanol extracts were effective on three kinds of citrus fungus pathogens, in which the best ECso was 0.289 0 mg·mL^（-1） against Coiletorichum gloeosporioides. Fraction B2 was analyzed to have four eoumarins including isobergapten, sphondin, pimpinellin and isopimpinellin. Antifungal virulence of isobergapten, pimpinellin and isopimpinellin against C. gloeosporioides show that EC50 were about 45 mg·L^（-1）, in which pimpinellin was 41.521 6 mg·L^（-1）. Pimpinellin could induce the mycelium branch increase, hypha deformity, partial septum disappear and so on. dentified to inhibit citrus [Conclusion]Isobergapten, pimpinellin and isopimpinellin were isolated and ifungus pathogens from Radix Angelicae Biseratae ethanol extracts.