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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
Four representative Fusarium graminearum isolates from China and seven from Europe were selected and identified by DON specific primers to confirm that they had the genetic material basis for producing DON toxin and then inoculated into PDB The culture medium was cultured for 7 days, and the DON toxin was purified from the supernatant of the medium by silica gel column. The quantitative analysis by HPLC showed that the purified DON toxin from Fusarium graminearum was the same as the standard DON poison sample purchased from the company. The HPLC detection spectrum The peak was clear and clear, the baseline was stable and without interference, the retention time was about 9.5min; the DON concentration of the tested strains ranged from 0.023μg / mL to 1.934μg / mL, and the German strain F703 produced the highest amount of toxin Strain 5005 (1.232μg / mL) was 57% higher than that of the other Chinese strains 7105 (0.135μg / mL), except for the Italian strain Lor9 (0.128μg / mL) Chinese strains (4020,7105,8029) produce large amount of toxin.Fusarium toxins produce DON toxin is far greater than the capacity of Chinese strains, indicating that European strains have long been in the European ecological environment has evolved its unique high-toxic metabolic types, It is necessary for our country to prevent Fusarium graminearum from entering Europe , To strengthen the quarantine and surveillance of Fusarium and toxins of cereal grains and their products from Europe.At the same time, the toxin sample preparation and HPLC detection system established in this study can be used to accurately quantify the DON toxins of the samples. 14