石楠拟盘多毛孢致病毒素的初步分离纯化

Separation and purification of toxins produced by Pestalotiopsis photiniae

为了获得引起草莓根腐病的石楠拟盘多毛孢(Pestalotiopsis photiniae)产生的致病毒素组分,利用中压硅胶柱层析、高效液相C-18制备柱层析和sephadex LH-20层析对粗毒素进行了分离纯化,以离体叶盘针刺法检测分离所得组分的致病活性。结果显示:经中压硅胶柱层析和高效制备液相色谱得到了FrⅢ1和FrⅢ2两个组分,其中FrⅢ1有较强活性;用sephadex LH-20对FrⅢ1进一步纯化后又得到了两组活性组分FrⅣ1-2和FrⅣ3-4,其中组分FrⅣ3-4的活性明显强于组分FrⅣ1-2;HPLC分析发现这两组组分又分别含有2个和5个组分,7个组分均在C-18柱层析过程中出峰时间较早,说明该毒素的极性较大。

In order to obtain the toxin components produced by Pestalotiopsis photiniae which causes the strawberry root rot,silica gel column chromatograph,C-18preparation-HPLC,and sephadex LH-20 column chromatograph were used for separating and purifying the toxin,the in vitro leaf disk acupuncture method was applied to detect the pathogenicity of separated toxin components.The results showed that the component FrⅢ1and FrⅢ2 were acquired through silica gel column chromatograph and preparation-HPLC,and FrⅢ1was more active than FrⅢ2on strawberry leaf disks.After sephadex LH-20 column chromatograph purification,two component groups named as FrⅣ1-2and FrⅣ3-4were gained from FrⅢ1,and the activity of FrⅣ3-4was stronger than that of FrⅣ1-2.HPLC analysis suggested that FrⅣ1-2and FrⅣ3-4contained 2and 5components,respectively.All the 7components showed short retention time through the C-18 column,which indicated that these fractions of the toxin had strong polarity.