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禾谷镰孢菌产玉米赤霉烯酮毒素zearalenone的PCR检测
引用本文:张瑞芳,马青,王晓鸣.禾谷镰孢菌产玉米赤霉烯酮毒素zearalenone的PCR检测[J].植物保护,2009,35(6):94-98.
作者姓名:张瑞芳  马青  王晓鸣
作者单位:1. 西北农林科技大学植物保护学院,陕西省农业分子生物学重点实验室,杨凌,712100;中国农业科学院作物科学研究所,国家农作物基因资源与基因改良重大科学工程,北京,100081
2. 西北农林科技大学植物保护学院,陕西省农业分子生物学重点实验室,杨凌,712100
3. 中国农业科学院作物科学研究所,国家农作物基因资源与基因改良重大科学工程,北京,100081
摘    要:玉米赤霉烯酮(zearalenone,ZEN)是由镰孢菌产生的真菌毒素,该毒素具有雌性激素生物活性。PKS4基因是禾谷镰孢菌中玉米赤霉烯酮生物合成途径中的必需基因。根据PKS4基因序列,设计一对PKS4基因特异性引物,通过检测PKS4基因的存在间接检测ZEN毒素的产生。特异引物在禾谷镰孢菌菌株以及被禾谷镰孢菌侵染的小麦籽粒中都能稳定地扩增出大小为1 076 bp的特异片段,扩增片段与PKS4基因(DQ019316)相同区段序列相似性达99.63%。同时,酶联免疫吸附法(ELISA)验证了PCR的检测结果,证明了该技术的可靠性。

关 键 词:禾谷镰孢菌  玉米赤霉烯酮  PCR  检测

Detection of zearalenone produced by Fusarium graminearum using PCR assay
Zhang Ruifang,Ma Qing,Wang Xiaoming.Detection of zearalenone produced by Fusarium graminearum using PCR assay[J].Plant Protection,2009,35(6):94-98.
Authors:Zhang Ruifang  Ma Qing  Wang Xiaoming
Institution:1. College of Plant Protection and Shaanxi Key Laboratory of Molecular Biology for Agriculture,Northwest Agricultural and Forestry University, Yangling712100, China;2. Institute of Crop Science / The National Key Facility for Crop Gene Resources and Genetic Improvement,NFCRI,Chinese Academy of Agricultural Sciences, Beijing100081, China
Abstract:Zearalenone(ZEN)is produced by some Fusarium species and is one of the estrogenic mycotoxins.The gene PKS4 is essential for ZEN synthesis in F.graminearum.A pair of primers was designed based on the nucleotide sequences of PKS4 gene and used for detecting ZEN indirectly.The primers can amplify a 1076 bp fragment not only in strains of F.graminearum but also in wheat seeds infected by F.graminearum.Sequence comparison indicared that the fragments amplified were 99.63% identity to the gene PKS4(DQ019316).The strains of F.graminearum and seeds producing ZEN identified by PCR were confirmed by ELISA testing.
Keywords:PCR  Fusarium graminearum  zearalenone  PCR  detection
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