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Areca palm velarivirus 1 (APV1) is identified as a causative agent of yellow leaf disease (YLD), which emerges as a prominent threat to betel palm plantation. Developing methods for rapid detection of APV1 is necessary for preventing and controlling YLD in betel palm cultivation. In this work, APV1 virions were extracted from APV1-infected Nicotiana benthamiana by using polyethylene glycol (PEG) precipitation, ultracentrifugation, sucrose density gradient centrifugation, and affinity magnetic beads. The purified APV1 virions were identified by transmission electron microscopy (TEM), SDS-PAGE and Western blotting. The purified APV1 virions were used to immunize BALB/c mice to produce polyclonal antiserum for detection of APV1. APV1 virions were precipitated from N. benthamiana homogenates by applying 5% PEG6000 and 0.6% NaCl. After resuspension and ultracentrifugation with 55% sucrose cushion, APV1 virions were distributed in the lower part of the sucrose layer or precipitated at the bottom of the centrifuge tube. APV1 was purified by 30%, 40%, 50%, 60% and 70% discontinuous sucrose density gradient centrifugation at 140 000×g for 2 h, and the results showed that APV1 was enriched in 60% and 70% sucrose layers. Affinity magnetic beads could efficiently purify APV1 virions. The purified virions were elongated, about 650–2200 nm in length and 10–13 nm in diameter. BALB/c mice were immunized with the extracted APV1 virions to obtain antiserum with high specificity for APV1 and titer of 1∶25 600. The results would provide a new idea for the separation and purification of APV1, and an important technical support for rapid detection of APV1.
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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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