应用多聚酶链式反应(PCR)技术检测和定量分析柑桔黄龙病病原

 应用多聚酶链式反应(PCR)技术对柑桔黄龙病病原进行检测和定量分析。研究结果表明:PCR技术能准确、快速地检测出田间和温室内罹病的柑桔、长春花以及带菌柑桔木虱样本中的柑桔黄龙病病原(BLO),而健康植株和柑桔木虱样本则呈阴性反应。采用的PCR引物(Primers)是根据柑桔黄龙病病原亚洲株系的DNA序列(In-2.6,基因库号码:M9491)设计的。应用竞争性定量多聚酶链式反应(Q-PCR)方法能测定病原在不同样品中的含量。该技术关键在于:病原DNA和浓度系列稀释后的竞争物DNA,在同一试管中进行DNA扩增反应时,共用相同的引物。由于彼此互相竞争反应引物的结果,病原的PCR产物和竞争物的PCR产物在数量上呈反向线性相关。因竞争物的浓度为已知,便可从线性关系中推算出病原的含量。

DETECTION AND QUANTITATION OF CITRUS HUANGLONGBING PATHOGEN BY POLYMERASE CHAIN REACTION

In this study, polymerase chain reaction (PCR) was developed for detection and quantification of the bacteria-like organism (BLO) that causes citrus Huang longbing/greening disease. The primers for the PCR were designed based on the sequence information of In-2.6 (Genebank accession number M 9491). By using the PCR procedure, the presence of citrus Huang longbing BLO could be detected in samples of infected citrus, periwinkle, and insect vector fed on the BLO-infected plants.