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小麦和玉米籽粒赤霉菌产毒类型的PCR检测

PCR Identification of Mycotoxin-chemotypes from Grains of Wheat and Maize Associated with Head Blight

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【作者】 武爱波李和平张静柏姚明镜许玉彬廖玉才

【Author】 WU Ai-bo~1, LI He-ping~1, ZHANG Jing-bai~1, YAO Ming-jing~1, XU Yu-bin~1, LIAO Yu-cai~(1,2)(1. Molecular Biotechnology Laboratory, Huazhong Agricultural University, Wuhan, Hubei 430070, China; 2. College of Plant Science and Technology, Huazhong Agricultural University, Wuhan, Hubei 430070, China)

【机构】 华中农业大学分子生物技术实验室华中农业大学分子生物技术实验室 湖北武汉430070湖北武汉430070湖北武汉430070华中农业大学植物科学技术学院

【摘要】 为了建立一种准确、快速鉴定赤霉菌毒素的方法,根据赤霉菌毒素生物合成调控路径T ri5-T ri6基因保守序列,设计了一对通用引物,用于检测和鉴别产生脱氧雪腐镰刀菌烯醇(DON)和雪腐镰刀菌烯醇(N IV)毒素的赤霉菌特异DNA片断。结果表明,产生毒素DON的赤霉菌具有一个300 bp的特异DNA片断,而产生毒素N IV的则为360 bp。PCR检测与HPLC或GC/M S化学分析结果完全一致。利用这一方法分析检测了小麦、玉米籽粒中赤霉菌产毒类型,发现除健康籽粒外,在所有轻微、中度、严重感染赤霉病的小麦或玉米籽粒中,均同时检测到DON及N IV的两种DNA片断,表明这一对通用引物对两种毒素特异DNA片断的扩增效率相同。这些结果说明,该技术是一种经济、快速、可靠的PCR检测技术,可广泛用于赤霉菌及其毒素检测鉴定中。

【Abstract】 To develop a rapid and accurate method for the identification of mycotoxin-chemtypes from Fusarium head blight (FHB), a pair of primers was designed based on the conserved sequences of Tri5-Tri6 genes involved in the biosynthesis of Fusarium mycotoxins, which could detect the DNA fragments specific for mycotoxin-chemotypes deoxynivalenol (DON) and nivalenol (NIV). The results indicated that a 300 bp fragment from DON-chemotypes and a 360 bp fragment from NIV-chemotypes were amplified. The mycotoxin-chemotypes identified by PCR were confirmed by chemical analysis of HPLC or GC/MS. Further analyses showed that both DON-and NIV-specific DNA fragments were simultaneously amplified from the grains of wheat and maize with a light, intermediate or severe symptom of FHB, whereas no PCR product was detected in healthy grains, indicating that the PCR assay is a cost-effective, rapid and reliable method, and could efficiently amplify fragments from two mycotoxin-chemotypes. This PCR assay could be widely used in detection of DON-and NIV-mycotoxin chemotypes for plant quarantine and the safety controls of food and livestock.

【关键词】 小麦玉米赤霉菌毒素检测PCR
【Key words】 WheatMaizeFusarium head blightMycotoxin identificationPCR
【基金】 国家自然科学基金项目(30470929);中-欧合作项目(ERBIC18CT980312)
  • 【文献出处】 麦类作物学报 ,Acta Tritical Crops , 编辑部邮箱 ,2005年05期
  • 【分类号】S432.4
  • 【被引频次】20
  • 【下载频次】451
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