Rapid and accurate diagnosis of fungal pathogens is essential for disease control in sunflower. In the present study, a multiplex PCR assay was developed based on the dual priming oligonucleotide (DPO) system, which was used to simultaneously detect and identify five major sunflower fungal pathogens. There was no cross-reactivity among the pathogens tested. In each reaction, 0.1 ng genomic DNA templates were sufficient to ensure specificity and accuracy. The system exhibited high adaptability over a wide range of annealing temperatures. No mismatch or nonspecific amplification was observed in the annealing temperatures tested. In combination with capillary electrophoresis, the DPO-primer-based multiplex PCR system provides a rapid, reliable and cost-efficient solution for the diagnosis of fungal pathogens in sunflower.

快速准确地诊断真菌病原体对于控制向日葵的病害至关重要。在本研究中，基于双引发寡核苷酸（DPO）系统开发了一种多重PCR检测方法，用于同时检测和鉴定五种主要的向日葵真菌病原体。测试的病原体之间没有交叉反应性。在每个反应中，0.1 ng 基因组 DNA 模板足以确保特异性和准确性。该系统在很宽的退火温度范围内表现出很高的适应性。在测试的退火温度中没有观察到错配或非特异性扩增。与毛细管电泳相结合，基于 DPO 引物的多重 PCR 系统为诊断向日葵中的真菌病原体提供了一种快速、可靠且具有成本效益的解决方案。