禾谷镰刀菌与亚细亚镰刀菌种型特异性的SCAR标记

摘要

为发掘区分Fusarium asiaticum与Fusarium graminearum这两种赤霉病菌的特异性分子标记,从江淮麦区赤霉病穗上分离获得72个赤霉病菌株,进行种型与化学型鉴定,并利用SRAP引物对F.asiaticum-NIV、F.asiaticum-3ADON及F.graminearum-15 ADON 3种类型的致病菌进行种型及化学型特异性标记筛选.通过对特异性标记进行回收、转化与测序,根据序列设计出3个镰刀菌种型特异性SCAR引物,并用上述72个菌株进行验证.结果显示SCAR标记me1/em2-172仅在F.asiaticum中扩增出172 bp的特异性条带,标记me1/em6-311和me9/em4-639分别只在F.graminearum中扩增出311 bp和639 bp的特异性条带.表明,本研究获得的3个SCAR标记是F.asiaticum和F.graminearum种型特异性标记,可用于区分这两种镰刀菌,为深入研究其致病机理及防治赤霉病的发生及危害提供了研究基础.%To develop specific markers for identifying the species and chemotypes of Fusarium asiaticum and Fusarium graminearum, infected spikes were collected from several wheat-growing regions between Yangtze River and Huaihe River, and seventy-two isolates collected from the diseased spikes were identified the genotype by primer pair Fgl6F/Fgl6R and chemotype by primer pair TOXP1/P2 and multiplex PCR. Sequence related amplified polymorphism (SRAP) markers were screened on representative isolates of F. asiaticum-NIV, F. asiaticum- 3ADON and F. graminearum-15ADON to find species-specific and chemotype-specific products. Three species-specific SRAP markers were obtained by cloning and sequencing the specific products. Sequence-characterized amplified region ( SCAR ) primers specific for F. asiaticum and F. graminearum were designed based on the sequences of these species-specific products. The SCAR primers were tested with 72 isolates. The results showed that the primer pair mel/em2-172 could amplify a 172 bp PCR product only in F. asiaticum isolates, while the primer pairs of mel/em6-311 and me9/em4-639 could amplify 311 bp and 639 bp PCR products only in F. graminearum rnisolates, respectively. These results indicated that the SCAR primers designed in this study were species-specific primers for F. asiaticum and F. graminearum and could be used to identify these two types of Fusarium. The information obtained in this study provided a better understanding of the pathogenicity of Fusarium spp. and references for disease management.