A Method to Inoculate Millet Grain-Colonized <em>Fusarium pseudograminearum</em> on Wheat to Obtain Reproducible Disease Symptoms

Fusarium crown rot (FCR), mainly caused by <i>Fusarium pseudograminearum</i>, is a devastating soil-borne disease of wheat that results in severe yield and quality reduction. FCR is characterized by stem base necrosis and whitehead development. In recent years, FCR has escalated in both incidence and severity, emerging as a critical threat to global wheat production, particularly within key cultivation zones such as China's Huang-Huai-Hai Plain. The development of resistant cultivars is an effective and environmentally sustainable strategy for FCR disease control. However, the lack of standardized and reproducible inoculation protocols has hindered the accurate assessment and screening of disease-resistant wheat germplasms. To address this limitation, we established a robust FCR inoculation system utilizing <i>F. pseudograminearum</i> propagated on a millet grain substrate, facilitating rapid and reliable evaluation of both host resistance and fungal pathogenicity. Laboratory validation demonstrated high infection efficiency and strong reproducibility of this method. Key features • Standardized and reproducible greenhouse inoculation protocol for Fusarium crown rot resistance assessment, established in wheat. • This system enables accurate quantification of <i>F. pseudograminearum</i> pathogenicity and facilitates high-throughput screening of resistant wheat germplasm.