中国与欧洲禾谷镰刀菌DON毒素HPLC定量比较分析

选用4个来自中国、7个来自欧洲的代表性禾谷镰刀菌菌株,经脱氧雪腐镰刀菌烯醇(DON)毒素特异引物鉴定,确认其具有产生DON毒素的遗传物质基础后,接种于PDB培养基培养7d,从培养基上清中经硅胶柱分离、纯化DON毒素,经HPLC定量分析表明,纯化的禾谷镰刀菌DON毒素,与购自公司的DON毒素标准样品一样,其HPLC检测谱峰清晰明显,基线平稳,无干扰,保留时间为9.5min左右;供试菌株的DON毒素含量分布在0.023~1.934μg/mL之间,德国菌株F703产毒量最大,比位居第二的中国菌株5005(1.232μg/mL)高57%;其余的6个欧洲菌株中,除意大利菌株Lor9(0.128μg/mL)略低于另一个中国菌株7105(0.135μg/mL)外,均比3个中国菌株(4020、7105、8029)的产毒量大.欧洲镰刀菌产生DON毒素的能力远远大于中国菌株,这说明欧洲菌株长期在欧洲生态环境下已演变形成其特有的高毒素代谢类型,我国有必要严防欧洲禾谷镰刀菌入侵,加强对来自欧洲的禾谷类粮食及其产品的镰刀菌和毒素的检疫和监控;同时,本研究建立的毒素样品制备与HPLC检测体系可用于准确定量分析样品的DON毒素.图3表2参14

Comparative and Quantitative Assessment of DON Levels of Fusarium graminearum Isolates from China and Europe by HPLC

Eleven Fusarium graminearum isolates,four from China and seven from Europe were first identified for their genetic basis for the synthesis of deoxynivalenol (DON) mycotoxins, with their identity confirmed as DON-producing isolates.The DON-producing F. graminearum isolates were then grown on PDB for 7 days, and DON mycotoxins were purified by silicon gel column and subjected to analysis by high performance liquid chromatography (HPLC). The results showed that the purified DON mycotoxins from F. graminearum displayed a clear, sharp peak without visible disturbance and had a retention time of about 9.5 min, which was almost identical to that obtained with standard DON mycotoxin samples purchased form a company. DON mycotoxin concentrations ranged from 0.023 to 1.934 g per milliliter. Among the eleven isolates, F703 from Germany produced the highest level of DON, an increase by 57 % compared to the highest producer of the Chinese isolates, 5005 (1.232 g/mL). The other six European isolates, except isolate Lor9 (0.128 g/mL) from Italy that produced slightly lower DON than isolate 7105 (0.135 g/mL) from China, contained much higher DON mycotoxins than the three Chinese isolates (4020, 7105, 8029). The higher DON mycotoxin-production capability of the European Fusarium isolates suggested that it was necessary to prevent invasion of F. graminearum isolates from Europe, and to strictly control and monitor DON mycotoxins and their producers from cereals or related food products imported from European countries. The present results also indicate that the methods described in this study for purifying DON mycotoxins, and the parameters and conditions established in the current HPLC assay may be useful for rapid and accurate analysis of DON mycotoxins. Fig 3, Tab 2, Ref 14