Russian State Agrarian University - Moscow Timiryazev Agricultural Academy, Moscow, Russia.
People's Friendship University of Russia - RUDN University, Moscow, Russia.
Braz J Biol. 2023 Oct 30;83:e275505. doi: 10.1590/1519-6984.275505. eCollection 2023.
Multiplex real-time PCR with TaqMan® probes has been developed for the simultaneous detection of soybean pathogens Pseudomonas savastanoi pv. glycinea and Curtobacterium flaccumfaciens pv. flaccumfaciens. The method specificity has been confirmed using 25 strains of target bacteria and 18 strains of other bacteria common to soybean seeds as endophytes. The multiplex real-time PCR developed has been shown to have high sensitivity - a positive result was achieved at 0.01 ng/µl of DNA for both target organisms, and at 100 CFU/ml of bacteria in soybean seed homogenate. The robustness of the multiplex real-time PCR developed has been verified by the detection of the pathogens in 25 commercial seed stocks, in comparison with previously published PCR protocols. In all tests, three seed stocks were positive and 22 were negative. The multiplex real-time PCR can be applied in diagnostic practice for the simultaneous detection of two important pathogens of leguminous plants.
已开发出一种用于同时检测大豆病原体丁香假单胞菌 pv. 大豆和黄单胞菌 pv. 叶斑病的多重实时 PCR 方法，使用 TaqMan®探针。该方法特异性已通过 25 株靶细菌和 18 株常见于大豆种子的内生细菌菌株得到证实。所开发的多重实时 PCR 具有高灵敏度-对于两种目标生物，在 DNA 浓度为 0.01 ng/µl 时即可获得阳性结果，在大豆种子匀浆中细菌浓度为 100 CFU/ml 时即可获得阳性结果。通过与先前发表的 PCR 方案比较，对 25 种商业种子库存中的病原体进行检测，验证了所开发的多重实时 PCR 的稳健性。在所有测试中，三种种子库存呈阳性，22 种呈阴性。该多重实时 PCR 可应用于诊断实践中，用于同时检测两种重要的豆科植物病原体。
