为了建立中国小麦秆锈菌流行小种的分子检测标记,利用RAPD(Random amplified polymorphic DNA)技术对我国小麦秆锈菌6个主要生理小种21C3CTH、21C3CPH、21C3CFH、34MKG、34C2MKK和34C2MKR进行了分析,筛选特异性片段并将其转化为SCAR(Sequence-characterized amplified region)标记。在156条10碱基随机引物中,引物S92(5′-CAGCTCACGA-3′)在小麦秆锈菌生理小种21C3CTH中扩增出一条782bp的特异片段,回收、克隆和测序该特异片段,根据特异片段的核苷酸序列设计了1对SCAR特异引物,经验证,该引物特异性良好。结果表明,小麦秆锈菌生理小种21C3CTH的RAPD标记被成功地转化为SCAR标记,这为该生理小种的分子鉴定和监测奠定了基础。
In order to establish the molecular markers to identify the prevalent races of Puccinia graminis f.sp.tritici(Pgt)in China,6 Pgt physiological races(21C3CTH,21C3CPH,21C3CFH,34MKG,34C2MKK and 34C2MKR)were analyzed using RAPD(Random amplified polymorphic DNA)technology,and the specific band were selected to develop SCAR(Sequence-characterized amplified region)marker.Out of 156RAPD primers,S92(5′-CAGCTCACGA-3′)amplified a 782bp specific fragment fromPgt race 21C3CTH.The band was purified,cloned in the pGM-T vector and sequenced.Based on the nucleotide sequence of the specific band,a new pair of PCR primers for SCAR marker of21C3CTH was designed using the Primer Premier 5.0software.It was validated that this marker was highly specific for 21C3CTH.The SCAR marker of Chinese Pgt race 21C3CTH developed was useful for the molecular identification and monitoring of 21C3CTH.