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Odontoglossum ringspot virus (ORSV), one of the major pathogens affecting orchids, seriously degrade the ornamental and economic value of orchids by causing leaf chlorosis, mottling, and floral deformities. To enable early-stage diagnosis, this study developed a rapid reverse transcription-recombinase polymerase amplification (RT-RPA) assay targeting ORSV. Primers and a fluorescent probe were designed based on conserved regions of the ORSV coat protein (CP) gene. Specificity, sensitivity and field applicability of the assay were systematically validated in this study. Results identified F1/R1 as the optimal primer pair, enabling amplification within 20 minutes at 41 ℃. The method exhibited high specificity, distinguishing ORSV from Cymbidium mosaic virus and Cucumber mosaic virus, and achieved a detection limit of 2.8×10–5 ng/μL total RNA. Practical verification result of 20 Phalaenopsis field samples showed 100% concordance with TaqMan qPCR results. The RT-RPA assay would provide a rapid, sensitive, and field-deployable tool for ORSV surveillance, supporting timely disease management and sustainable orchid cultivation. Its simplicity and reliability make it particularly valuable for large-scale screening in industrial orchid production systems.
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| 科 Family | 属数 Number of genus | 种数 Number of species | 占总种数比例 Percentage of total species (%) | 属 Genus | 种数 Number of species | 占总种数比例 Percentage of total species (%) |
|---|---|---|---|---|---|---|
| 鹅膏菌科Amanitaceae | 2 | 11 | 5.26 | 鹅膏菌属 Amanita | 10 | 4.78 |
| 小菇科 Mycenaceae | 2 | 12 | 5.74 | 丝盖伞属 Inocybe | 5 | 2.39 |
| 多孔菌科 Polyporaceae | 8 | 14 | 6.70 | 蜡蘑属 Laccaria | 5 | 2.39 |
| 红菇科 Russulaceae | 3 | 23 | 11.00 | 小皮伞属 Marasmius | 6 | 2.87 |
| 小菇属 Mycena | 11 | 5.26 | ||||
| 光柄菇属 Pluteus | 5 | 2.39 | ||||
| 红菇属 Russula | 17 | 8.13 | ||||
| 栓菌属 Trametes | 5 | 2.39 |
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