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| Rapid Identification of Main Pathogen Fusarium pseudograminearum of Wheat Crown Rot Using Site-specific PCR Based on EF-1α Sequence |
| LIU Guo-Xia1,*, TAN Qing-Qing1, QI Jun-Shan2, WANG Fu-Yu3, CHEN Xue-Yan1, FAN Yang-Yang1, HU Yue1, BU Xun1, ZHANG Quan-Fang1,* |
1 Biotechnology Research Center, Shandong Academy of Agricultural Sciences, Jinan 250100, China;
2 Institute of Plant Protection, Shandong Academy of Agricultural Sciences, Jinan 250100, China;
3 Jining Academy of Agricultural Sciences, Jining 272031, China |
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Abstract Wheat stem rot (WCR) is an important disease affecting wheat yield, Fusarium pseudograminearum is the main pathogen. The rapid and accurate identification of pathogens is significant for making reasonable control measures and resistant varieties breeding. In this study, elongation factor 1α (EF-1α) gene sequences of common Fusarium spp. were compared and specific primers F.pseudo-F3/R1 for F. pseudograminearum were designed, touchdown PCR were used to specifically amplify a 334 bp band from F. pseudograminearum, and no band amplified in F. graminearum, F. tricinctum, F. proliferatum, F. incarnatum and F. oxysporum. Wheat samples collected from Jining and Jiyang in Shandong province in 2019 and 2020 were detected using the site-specific PCR method which was just developed to test its validity. It was found that 28 fungi strains were isolated from infected samples from Jining in 2019, in which F. pseudograminearum was amplified with aimed band of 334 bp using primers F.pseudo-F3/R1, while no band from other fungi including F. graminearum, F. equiseti, Alternaria alternate, Bipolaris sorokiniana and Rhizoctonia cerealis. Samples were also collected from Jining in 2020 and DNA isolated directly from wheat stem with symptom of crown rot, then amplified using primers F.pseudo-F3/R1 and got a 334 bp aimed band of F. pseudograminearum, while other samples with symptom of root rot, leaf rust, powdery mildew or white-headed got no amplification band. The fungi isolated from stem of samples used above from Jining in 2020 were identified by sequencing analysis, and the result showed that F. pseudograminearum and F. graminearum existed in crown rot wheat sample, while F. graminearum and R.cerealis existed in other samples. The wheat stems of artificially and naturally infected by F. pseudograminearum from Jiyang in 2020 were amplified using primers F.pseudo-F3/R1 and got a 334 bp aimed band of F. pseudograminearum. The above results indicated that the specific PCR method could be used to detect F. pseudograminearum in stem directly, which provides a new method to rapidly identify F. pseudograminearum.
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Received: 23 October 2020
Published: 01 May 2021
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Corresponding Authors:
*liu_guo_xia@163.com;zhquanfang@163.com
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