Phytopathology. 2005 Dec;95(12):1423-9. doi: 10.1094/PHYTO-95-1423.
ABSTRACT Reliable and sensitive quantification of Phytophthora capsici in pepper plants is of crucial importance in managing the multiple syndromes caused by this pathogen. A real-time polymerase chain reaction (PCR) assay was developed for the determination of P. capsici in pepper tissues. DNA levels of a highly virulent and a less virulent isolate were measured in different pepper genotypes with varying degrees of resistance. Using SYBR Green and specific primers for P. capsici, the minimal amount of pathogen DNA quantified was 10 pg. Pathogen DNA was recorded as early as 8 h postinoculation. Thereafter, the increase was rapid in susceptible cultivars and slower in resistant ones. The amount of pathogen DNA quantified in each pepper genotype correlated with susceptibility to Phytophthora root rot. Likewise, there was a relationship between the virulence of the pathogen and the degree of colonization. Differences also were found in oomycete amount among pepper tissues, with maximal pathogen biomass occurring in stems. The real-time PCR technique developed in this study was sensitive and robust enough to assess both pathogen development and resistance to Phytophthora root rot in different pepper genotypes.
在管理由该病原体引起的多种综合征时,可靠且灵敏地定量检测辣椒疫霉在辣椒植株中的含量至关重要。本研究开发了一种用于检测辣椒组织中辣椒疫霉的实时聚合酶链反应(PCR)检测方法。使用 SYBR Green 和针对辣椒疫霉的特异性引物,该方法能够检测到最低 10 pg 的病原菌 DNA。在接种后 8 小时即可检测到病原菌 DNA,此后,在感病品种中病原菌 DNA 的增加迅速,而在抗病品种中则较慢。在每个辣椒品种中定量检测到的病原菌 DNA 与对辣椒疫霉根腐病的敏感性相关。同样,病原菌的毒力与其定殖程度之间也存在相关性。在不同的辣椒组织中,疫霉菌的含量也存在差异,其中茎中的病原菌生物量最大。本研究中开发的实时 PCR 技术灵敏且稳健,足以评估不同辣椒品种中病原菌的生长和对辣椒疫霉根腐病的抗性。