Development and validation of a rapid loop-mediated isothermal amplification assay for the detection of Chrysomyxa and characterization of Chrysomyxa woroninii overwintering on Picea in China
Abstract Chrysomyxa rusts cause significant damage to spruce in both natural forests and plantations. Particularly, Three Chrysomyxa species, Chrysomyxa deformans, Chrysomyxa qilianensis, and Chrysomyxa rhododendri, listed as National Forest Dangerous Pests in China, have severely affected many econ...
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BMC
2024-08-01
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| Series: | IMA Fungus |
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| Online Access: | https://doi.org/10.1186/s43008-024-00157-6 |
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| author | Wan Ting Yu Xin Wang Tan Yin Clement Kin-Ming Tsui Chong Juan You |
| author_facet | Wan Ting Yu Xin Wang Tan Yin Clement Kin-Ming Tsui Chong Juan You |
| author_sort | Wan Ting Yu |
| collection | DOAJ |
| description | Abstract Chrysomyxa rusts cause significant damage to spruce in both natural forests and plantations. Particularly, Three Chrysomyxa species, Chrysomyxa deformans, Chrysomyxa qilianensis, and Chrysomyxa rhododendri, listed as National Forest Dangerous Pests in China, have severely affected many economically and ecologically important spruce native species in China. Also, Chrysomyxa arctostaphyli, an important plant quarantine fungus, causes a damaging broom rust disease on spruce. Therefore, rapid, and efficient detection tools are urgently needed for proper rust disease detection and management. In this study, a sensitive, genus-specific loop-mediated isothermal amplification (LAMP) assay targeting the ITS-28S rRNA region was developed to detect the presence of Chrysomyxa in spruce needle and bud samples. After optimization and validation, the LAMP assay was found to be sensitive to detect as low as 5.2 fg/µL DNA, making it suitable for rapid on-site testing for rust infection. The assay was also specific to Chrysomyxa species, with no positive signals from other rust genus/species. The application of LAMP in the early detection of rust infections in spruce needles and buds was investigated, and spatial colonization profiles as well as the means of overwintering of Chrysomyxa woroninii in infected buds and branches were verified using the LAMP assay. This LAMP detection method will facilitate further studies on the characteristics of the life cycle and inoculation of other systemic rusts. |
| format | Article |
| id | doaj-art-47b7a346403d429ea5740586989cee21 |
| institution | Kabale University |
| issn | 2210-6359 |
| language | English |
| publishDate | 2024-08-01 |
| publisher | BMC |
| record_format | Article |
| series | IMA Fungus |
| spelling | doaj-art-47b7a346403d429ea5740586989cee212025-02-03T09:40:41ZengBMCIMA Fungus2210-63592024-08-0115111510.1186/s43008-024-00157-6Development and validation of a rapid loop-mediated isothermal amplification assay for the detection of Chrysomyxa and characterization of Chrysomyxa woroninii overwintering on Picea in ChinaWan Ting Yu0Xin Wang1Tan Yin2Clement Kin-Ming Tsui3Chong Juan You4Beijing Key Laboratory for Forest Pest Control, Beijing Forestry UniversityBeijing Key Laboratory for Forest Pest Control, Beijing Forestry UniversityBeijing Key Laboratory for Forest Pest Control, Beijing Forestry UniversityInfectious Disease Research Laboratory, National Centre for Infectious Diseases, Tan Tock Seng HospitalBeijing Key Laboratory for Forest Pest Control, Beijing Forestry UniversityAbstract Chrysomyxa rusts cause significant damage to spruce in both natural forests and plantations. Particularly, Three Chrysomyxa species, Chrysomyxa deformans, Chrysomyxa qilianensis, and Chrysomyxa rhododendri, listed as National Forest Dangerous Pests in China, have severely affected many economically and ecologically important spruce native species in China. Also, Chrysomyxa arctostaphyli, an important plant quarantine fungus, causes a damaging broom rust disease on spruce. Therefore, rapid, and efficient detection tools are urgently needed for proper rust disease detection and management. In this study, a sensitive, genus-specific loop-mediated isothermal amplification (LAMP) assay targeting the ITS-28S rRNA region was developed to detect the presence of Chrysomyxa in spruce needle and bud samples. After optimization and validation, the LAMP assay was found to be sensitive to detect as low as 5.2 fg/µL DNA, making it suitable for rapid on-site testing for rust infection. The assay was also specific to Chrysomyxa species, with no positive signals from other rust genus/species. The application of LAMP in the early detection of rust infections in spruce needles and buds was investigated, and spatial colonization profiles as well as the means of overwintering of Chrysomyxa woroninii in infected buds and branches were verified using the LAMP assay. This LAMP detection method will facilitate further studies on the characteristics of the life cycle and inoculation of other systemic rusts.https://doi.org/10.1186/s43008-024-00157-6DNA-based diagnosticsChrysomyxaLAMPSpruce needle rustBud rust |
| spellingShingle | Wan Ting Yu Xin Wang Tan Yin Clement Kin-Ming Tsui Chong Juan You Development and validation of a rapid loop-mediated isothermal amplification assay for the detection of Chrysomyxa and characterization of Chrysomyxa woroninii overwintering on Picea in China IMA Fungus DNA-based diagnostics Chrysomyxa LAMP Spruce needle rust Bud rust |
| title | Development and validation of a rapid loop-mediated isothermal amplification assay for the detection of Chrysomyxa and characterization of Chrysomyxa woroninii overwintering on Picea in China |
| title_full | Development and validation of a rapid loop-mediated isothermal amplification assay for the detection of Chrysomyxa and characterization of Chrysomyxa woroninii overwintering on Picea in China |
| title_fullStr | Development and validation of a rapid loop-mediated isothermal amplification assay for the detection of Chrysomyxa and characterization of Chrysomyxa woroninii overwintering on Picea in China |
| title_full_unstemmed | Development and validation of a rapid loop-mediated isothermal amplification assay for the detection of Chrysomyxa and characterization of Chrysomyxa woroninii overwintering on Picea in China |
| title_short | Development and validation of a rapid loop-mediated isothermal amplification assay for the detection of Chrysomyxa and characterization of Chrysomyxa woroninii overwintering on Picea in China |
| title_sort | development and validation of a rapid loop mediated isothermal amplification assay for the detection of chrysomyxa and characterization of chrysomyxa woroninii overwintering on picea in china |
| topic | DNA-based diagnostics Chrysomyxa LAMP Spruce needle rust Bud rust |
| url | https://doi.org/10.1186/s43008-024-00157-6 |
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