Plant Breeding Institute, Christian-Albrechts-University of Kiel, Olshausenstr. 40, 24098, Kiel, Germany.
Sci Rep. 2023 Aug 3;13(1):12602. doi: 10.1038/s41598-023-39559-8.
Root lesion nematodes (RLN) of the genus Pratylenchus are causing significant damage in cereal production worldwide. Due to climate change and without efficient and environment-friendly treatments, the damages through RLNs are predicted to increase. Microscopic assessments of RLNs in the field and the greenhouses are time-consuming and laborious. As a result, cereal breeders have mostly ignored this pest. We present a method measuring RLN in infected cereal roots using a standardized PCR approach. Publicly available Pratylenchus neglectus primer combinations were evaluated. An optimal primer combination for RT-qPCR assay was identified to detect and quantify P. neglectus within infected cereal roots. Using the RT-qPCR detection assay, P. neglectus could be clearly distinguished from other plant parasitic nematodes. We could identify P. neglectus DNA in barley and wheat roots as low as 0.863 and 0.916 ng/µl of total DNA, respectively. A single P. neglectus individual was detected in water suspension and within barley and wheat roots. The RT-qPCR detection assay provides a robust and accurate alternative to microscopic nematode identification and quantification. It could be of interest for resistance breeding, where large populations must be screened to detect and quantify P. neglectus in farmer's fields.
根结线虫（RLN）属的普拉提尼恩斯（Pratylenchus）在全球范围内对谷物生产造成了重大损害。由于气候变化和缺乏有效且环保的处理方法，预计通过 RLN 造成的损害将会增加。在田间和温室中对 RLN 进行微观评估既耗时又费力。因此，谷物培育者大多忽略了这一害虫。我们提出了一种使用标准化 PCR 方法测量感染谷物根系中 RLN 的方法。评估了公开可用的普拉提尼恩斯忽视（Pratylenchus neglectus）引物组合。确定了用于 RT-qPCR 分析的最佳引物组合，以检测和量化感染谷物根系中的 P. neglectus。使用 RT-qPCR 检测分析，可以清楚地区分 P. neglectus 与其他植物寄生线虫。我们可以在大麦和小麦根系中低至 0.863 和 0.916 ng/µl 总 DNA 的水平上识别出 P. neglectus DNA。在水悬浮液以及大麦和小麦根系中均可检测到单个 P. neglectus 个体。RT-qPCR 检测分析为微观线虫鉴定和量化提供了一种强大而准确的替代方法。对于抗性育种来说，这可能很有趣，因为需要筛选大量种群以在农民的田地中检测和量化 P. neglectus。
