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| Establishment and Application of Dual PCR Method for Detection of Puccinia polysora and P. sorghi |
| LIN Fan-Kang1,2, WANG Shuo1, GUO Cheng3, GONG Xiang1, NIU Wen-Long1, SHI Yan1, LI Hong-Lian1,2, LI Yu1,2,*, WANG Ke1,* |
1 College of Plant Protection, Henan Agricultural University, Zhengzhou 450046, China;
2 State Key Laboratory of High-Efficiency Production of Wheat-Maize Double Cropping, Henan Agricultural University, Zhengzhou 450046, China;
3 Institute of Plant Protection, Gansu Academy of Agricultural Sciences, Lanzhou 730070, China |
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Abstract Rust is an important fungal disease in maize (Zea mays) production. In China, maize rust mainly includes southern corn rust caused by Puccinia polysora and common rust caused by P. sorghi. At present, relevant studies remain relatively limited on molecular detection technology of corn rust. Common detection methods have low sensitivity and are prone to false positives. In this study, the specific primers were designed and the amplification conditions were optimized to improve the specificity and sensitivity of the pathogen DNA detection by using the internal transcribed spacer (ITS) sequence of the P. polysora and P. sorghi as the target gene. A dual PCR detection method for these 2 rust fungi was established and applied to the detection of field samples. The results showed that the optimal reaction conditions of the dual PCR system were as follows: The final amount of primers was 1 μL (10 μmol/L), the annealing temperature was 55 ℃, the annealing time was 60 s, and the extension time was 90 s. The dual PCR system established in this study could detect the DNA of P. polysora and P. sorghi at a minimum concentration of 0.2 pg/μL, and its detection sensitivity was higher than that of conventional PCR methods. The detection results of 50 field samples showed that 38 samples were infected by P. polysora, 9 samples were infected by P. sorghi, and 3 samples were infected by P. polysora and P. sorghi. The dual PCR system established in this study can be used for rapid, sensitive and accurate detection of P. polysora and P. sorghi in the early stage, which provides technical support for early warning and later control of corn rust.
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Received: 15 September 2025
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Corresponding Authors:
*scauwangke@126.com; liyuzhibao@henau.edu.cn
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