Xu Jieying, Yang Xiao, Wu Cuiping, Chen Zhenpeng, Dai Tingting
Co-Innovation Center for the Sustainable Forestry in Southern China, Nanjing Forestry University, Nanjing, China.
Plant and Pest Diagnostic Clinic, Department of Plant Industry, Clemson University, Pendleton, SC, U.S.A.
Plant Dis. 2023 Apr;107(4):1067-1074. doi: 10.1094/PDIS-04-22-0864-RE. Epub 2023 Apr 24.
Pitch canker caused by the fungus is an important disease affecting pine trees in Europe and South Africa. Several countries, including China, have listed as a quarantine pathogen. Therefore, timely detection of could efficiently prevent its introduction into new areas or facilitate spread management in already infected sites. In this study, a recombinase polymerase amplification-lateral flow dipstick (RPA-LFD) assay was developed for rapid detection of based on a new target gene, , identified from whole-genome sequences. The assay was highly specific to . In fact, it exclusively detected isolates; 53 isolates of fungal and oomycete species and 2 nematodes of and were not detected. By detecting as little as 10 pg of genomic DNA in a 50-µl reaction, the RPA-LFD assay was 10 times more sensitive than conventional PCR assays. was also detected in artificially inoculated pine needles of . These results demonstrated that the developed RPA-LFD assay has the potential for rapid detection of in regions at high risk of infection. The RPA-LFD assay might serve as an alternative method for the early detection of .
由该真菌引起的松材线虫病是影响欧洲和南非松树的一种重要病害。包括中国在内的几个国家已将其列为检疫性病原菌。因此，及时检测该病原菌可有效防止其传入新的地区，或有助于在已感染地区进行传播管理。在本研究中，基于从全基因组序列中鉴定出的一个新的靶基因，开发了一种重组酶聚合酶扩增-侧向流动试纸条（RPA-LFD）检测方法，用于快速检测该病原菌。该检测方法对该病原菌具有高度特异性。事实上，它仅能检测到该病原菌的分离株；未检测到53株真菌和卵菌物种以及2种松材线虫和腐烂茎线虫的线虫。通过在50 μl反应中检测低至10 pg的该病原菌基因组DNA，RPA-LFD检测方法的灵敏度比传统PCR检测方法高10倍。在人工接种的该病原菌松针中也检测到了该病原菌。这些结果表明，所开发的RPA-LFD检测方法具有在高感染风险地区快速检测该病原菌的潜力。RPA-LFD检测方法可能成为该病原菌早期检测的一种替代方法。
