刺苋和苋属的实时荧光PCR鉴定

Real-time PCR Identification of Amaranthus spinosus and Amaranthus spp.

基于叶绿体的Psb A基因(Genbank登录号:DQ006133.1),设计了ASP-1和ASP-2两组具有刺苋特异性的引物探针,对苋科Amaranthaceae苋属Amaranthus刺苋A.spinosus的9个不同地理种、长芒苋A.palmeri的2个不同地理种、其它5种苋属植物和苋科非苋属8种,以及苋科外植物3种共27个样品进行Taq Man实时荧光PCR鉴定。结果表明,ASP-1适合作为刺苋的实时荧光PCR鉴定引物,其最佳扩增温度62℃;ASP-2适合作为苋属的实时荧光PCR鉴定,其最佳扩增温度为60℃。首次建立了Taq Man探针实时荧光PCR快速鉴定刺苋和苋属植物的方法。在优化后的检测条件下,ASP-1的检测灵敏度可达到0.03 ng·μL^(-1);ASP-2的检测灵敏度则可达到0.01 ng·μL^(-1)。上述方法作为形态鉴定的辅助方法,可准确有效的对刺苋及苋属植物进行鉴定。

Based on the conserved fragment of plant chloroplast genes（Genbank No.DQ006133.1）,a primer-probe set specific for Amaranthus spinosus（ASP-1）and a primer-probe set specific for Amaranthus spp.（ASP-2）were designed.Screened by to the 27 species including 9 target species,7 closely-related species and 11 other non-target species,the two primer-probe sets show good specificity.According to the real-time PCR tests results,the optimum amplification temperatures for ASP-1 and ASP-2 are 62℃and 60℃separately.Under optimal conditions ASP-1 and ASP-2 showed high efficiency in the detection of target plants with the detection limits of 0.03 ng·μL^（-1） for A.spinosus and 0.01 ng·μL^（-1） for Amaranthus spp.,respectively.It is the first time to establish a real-time PCR method for rapid identification of Amaranthus spinosus and Amaranthus spp.,and as an auxiliary method of morphological identification,it can be implemented accurately and effectively.