Establishment of recombinase polymerase amplification detection method for Dactylobotrys graminicola
Abstract Hulless barley sheath rot is a spike disease caused by Dactylobotrys graminicola. In recent years, it has generally occurred in hulless barley growing areas in China, resulting in reduced hulless barley yields. In this study, primers and probes were designed based on conserved genome sequen...
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Nature Portfolio
2024-10-01
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| Series: | Scientific Reports |
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| Online Access: | https://doi.org/10.1038/s41598-024-76921-w |
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| author | Haiqing Zhang Liyifan Chen Ruifang Dong Haowen Zheng Lu Hou Qiang Yao |
| author_facet | Haiqing Zhang Liyifan Chen Ruifang Dong Haowen Zheng Lu Hou Qiang Yao |
| author_sort | Haiqing Zhang |
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| description | Abstract Hulless barley sheath rot is a spike disease caused by Dactylobotrys graminicola. In recent years, it has generally occurred in hulless barley growing areas in China, resulting in reduced hulless barley yields. In this study, primers and probes were designed based on conserved genome sequences, and a method was established using recombinant enzyme polymerase amplification-lateral flow burette (RPA-LFD) technology for the rapid diagnosis of sheath rot in hulless barley. The method can be successfully established in five minutes at a constant temperature of 39℃, and the results are consistent with those of normal PCR analysis. The method demonstrated high sensitivity, with a detection limit of 10 fg/µL. Furthermore, the rapid method was able to successfully detect D. graminicola in hulless barley during field incubation, which highlighted the significant advantage of the method in practical applications. In conclusion, the RPA method established in this study exhibited several advantageous characteristics, including high efficiency, simplicity, rapidity and practicality, which provide a theoretical basis for the early detection and prevention of D. graminicola. |
| format | Article |
| id | doaj-art-b9de2e58109048829a372e080bc0f146 |
| institution | OA Journals |
| issn | 2045-2322 |
| language | English |
| publishDate | 2024-10-01 |
| publisher | Nature Portfolio |
| record_format | Article |
| series | Scientific Reports |
| spelling | doaj-art-b9de2e58109048829a372e080bc0f1462025-08-20T02:11:18ZengNature PortfolioScientific Reports2045-23222024-10-0114111110.1038/s41598-024-76921-wEstablishment of recombinase polymerase amplification detection method for Dactylobotrys graminicolaHaiqing Zhang0Liyifan Chen1Ruifang Dong2Haowen Zheng3Lu Hou4Qiang Yao5Qinghai Provincial Key Laboratory of Agricultural Integrated Pest Management, Academy of Agriculture and Forestry Science, Qinghai UniversityQinghai Provincial Key Laboratory of Agricultural Integrated Pest Management, Academy of Agriculture and Forestry Science, Qinghai UniversityQinghai Provincial Key Laboratory of Agricultural Integrated Pest Management, Academy of Agriculture and Forestry Science, Qinghai UniversityQinghai Provincial Key Laboratory of Agricultural Integrated Pest Management, Academy of Agriculture and Forestry Science, Qinghai UniversityQinghai Provincial Key Laboratory of Agricultural Integrated Pest Management, Academy of Agriculture and Forestry Science, Qinghai UniversityQinghai Provincial Key Laboratory of Agricultural Integrated Pest Management, Academy of Agriculture and Forestry Science, Qinghai UniversityAbstract Hulless barley sheath rot is a spike disease caused by Dactylobotrys graminicola. In recent years, it has generally occurred in hulless barley growing areas in China, resulting in reduced hulless barley yields. In this study, primers and probes were designed based on conserved genome sequences, and a method was established using recombinant enzyme polymerase amplification-lateral flow burette (RPA-LFD) technology for the rapid diagnosis of sheath rot in hulless barley. The method can be successfully established in five minutes at a constant temperature of 39℃, and the results are consistent with those of normal PCR analysis. The method demonstrated high sensitivity, with a detection limit of 10 fg/µL. Furthermore, the rapid method was able to successfully detect D. graminicola in hulless barley during field incubation, which highlighted the significant advantage of the method in practical applications. In conclusion, the RPA method established in this study exhibited several advantageous characteristics, including high efficiency, simplicity, rapidity and practicality, which provide a theoretical basis for the early detection and prevention of D. graminicola.https://doi.org/10.1038/s41598-024-76921-wHulless barley sheath rot diseaseDactylobotrys graminicolaRecombinase polymerase amplification lateral flow dipstick (RPA-LFD) |
| spellingShingle | Haiqing Zhang Liyifan Chen Ruifang Dong Haowen Zheng Lu Hou Qiang Yao Establishment of recombinase polymerase amplification detection method for Dactylobotrys graminicola Scientific Reports Hulless barley sheath rot disease Dactylobotrys graminicola Recombinase polymerase amplification lateral flow dipstick (RPA-LFD) |
| title | Establishment of recombinase polymerase amplification detection method for Dactylobotrys graminicola |
| title_full | Establishment of recombinase polymerase amplification detection method for Dactylobotrys graminicola |
| title_fullStr | Establishment of recombinase polymerase amplification detection method for Dactylobotrys graminicola |
| title_full_unstemmed | Establishment of recombinase polymerase amplification detection method for Dactylobotrys graminicola |
| title_short | Establishment of recombinase polymerase amplification detection method for Dactylobotrys graminicola |
| title_sort | establishment of recombinase polymerase amplification detection method for dactylobotrys graminicola |
| topic | Hulless barley sheath rot disease Dactylobotrys graminicola Recombinase polymerase amplification lateral flow dipstick (RPA-LFD) |
| url | https://doi.org/10.1038/s41598-024-76921-w |
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