Centro de Investigação de Montanha (CIMO), Instituto Politécnico de Bragança, Campus de Santa Apolónia, 5300-253, Bragança, Portugal.
Departamento de Ingeniería Agrária, Universidad de Léon, Av. Portugal, n° 41, 24071, Léon, Spain.
Sci Rep. 2022 Jan 11;12(1):492. doi: 10.1038/s41598-021-04422-1.
Philaenus spumarius is a vector of Xylella fastidiosa, one of the most dangerous plants pathogenic bacteria worldwide. There is currently no control measure against this pathogen. Thus, the development of vector control strategies, like generalist predators, such as spiders, could be essential to limit the spread of this vector-borne pathogen. In this study, a polymerase chain reaction (PCR)-based approach was developed to principally detect DNA of P. spumarius in the spider's gut. Accordingly, 20 primer pairs, targeting the mitochondrial cytochrome oxidase I (COI) and cytochrome b (cytB) genes, were tested for specificity, sensitivity, and efficiency in detecting P. spumarius DNA. Overall, two primer sets, targeting COI gene (COI_Ph71F/COI_Ph941R) and the cytB gene (cytB_Ph85F/cytB_Ph635R), showed the highest specificity and sensitivity, being able to amplify 870 pb and 550 bp fragments, respectively, with P. spumarius DNA concentrations 100-fold lower than that of the DNA of non-target species. Among these two primer sets, the cytB_Ph85F/cytB_Ph635R was able to detect P. spumarius in the spider Xysticus acerbus, reaching 50% detection success 82 h after feeding. The feasibility of this primer set to detect predation of P. spumarius by spiders was confirmed in the field, where 20% of the collected spiders presented positive amplifications.
沫蝉 Philaenus spumarius 是 Xylella fastidiosa 的传播媒介，Xylella fastidiosa 是世界上最危险的植物病原菌之一。目前针对这种病原体还没有控制措施。因此，开发传播媒介控制策略，如以沫蝉为食的捕食性蜘蛛等，对于限制这种媒介传播病原体的传播可能至关重要。在这项研究中，开发了一种基于聚合酶链反应 (PCR) 的方法，主要用于检测蜘蛛肠道中沫蝉的 DNA。为此，测试了 20 对针对线粒体细胞色素氧化酶 I (COI) 和细胞色素 b (cytB) 基因的引物对，以评估其检测沫蝉 DNA 的特异性、灵敏度和效率。总体而言，针对 COI 基因 (COI_Ph71F/COI_Ph941R) 和 cytB 基因 (cytB_Ph85F/cytB_Ph635R) 的两个引物对显示出最高的特异性和灵敏度，能够分别扩增 870 pb 和 550 pb 片段，而沫蝉 DNA 浓度比非目标物种的 DNA 低 100 倍。在这两个引物对中，cytB_Ph85F/cytB_Ph635R 能够在捕食性蜘蛛 Xysticus acerbus 中检测到沫蝉，在取食后 82 小时达到 50%的检测成功率。该引物对在野外检测沫蝉被蜘蛛捕食的可行性得到了证实，在采集的蜘蛛中有 20%呈现阳性扩增。
