Institute for Plant Protection, National Agriculture and Food Research Organization (NARO), Akitsu, Higashihiroshima, Hiroshima 739-2494, Japan.
Institute for Plant Protection, National Agriculture and Food Research Organization (NARO), Akitsu, Higashihiroshima, Hiroshima 739-2494, Japan.
J Microbiol Methods. 2021 Oct;189:106321. doi: 10.1016/j.mimet.2021.106321. Epub 2021 Sep 4.
Xylella fastidiosa causes many economically important plant diseases such as Pierce's disease of grapevine, citrus variegated chlorosis disease, and olive quick decline syndrome. Another species in the same genus, Xylella taiwanensis, causes pear leaf scorch. Here, to enable an initial screening of plants suspected of being infected with Xylella spp. by conventional polymerase chain reaction (cPCR), new primer pairs-X67S1/XL2r and XrDf1/XLr4-were designed to target the 16S ribosomal DNA (rDNA) of not only X. fastidosa but also X. taiwanensis. In cPCR to detect both species, X67S1/XL2r showed features superior to those of other primer pairs, such as fewer false negatives and false positives, whereas XrDf1/XLr4 seemed to be unsuitable because of abundant non-specific amplification. However, when XrDf1/XLr4 was combined with a probe in a TaqMan quantitative real-time PCR (qPCR), the assay detected no false positives and was more useful in the universal detection of Xylella spp. than TaqMan qPCR assays reported previously.
韧皮部杆菌属的 Xylella fastidiosa 可引起许多重要的植物病害，如葡萄藤的皮尔氏病、柑橘斑驳黄化病和油橄榄快速衰退综合征。该属的另一个种，即 Xylella taiwanensis，可引起梨叶枯焦病。在这里，为了能够通过常规聚合酶链反应（cPCR）对疑似感染韧皮部杆菌属的植物进行初步筛选，设计了新的引物对 X67S1/XL2r 和 XrDf1/XLr4，不仅可以针对 X. fastidosa，还可以针对 X. taiwanensis 的 16S 核糖体 DNA（rDNA）。在用于检测两个种的 cPCR 中，X67S1/XL2r 显示出优于其他引物对的特征，例如假阴性和假阳性较少，而 XrDf1/XLr4 似乎不合适，因为存在大量非特异性扩增。然而，当 XrDf1/XLr4 与 TaqMan 实时定量 PCR（qPCR）中的探针结合使用时，该检测方法没有假阳性，并且比之前报道的 TaqMan qPCR 检测方法更有助于通用检测 Xylella spp.。
