禾谷镰孢菌产玉米赤霉烯酮毒素zearalenone的PCR检测

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.PKS4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PKS4基因序列,设计一对PKS4基因特异性引物,通过检测PKS4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PKS4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.%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.