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In China, areca palm yellow leaf disease (AYLD) is primarily attributed to infection with phytoplasmas from the 16SrI, 16SrII and 16SrXXXII groups. Currently, detection methods predominantly target the 16SrI group or non-specifically detect all three groups, lacking specific identification methods for the 16SrII group phytoplasmas. To achieve specific detection of 16SrII group phytoplasmas, specific nested PCR primers were designed and screened in this study based on the ribosomal protein (rp) gene sequence of 16SrII group phytoplasmas, and the characteristics and application efficacy of this method were subsequently evaluated. Results showed that the method could accurately distinguish positive samples from healthy ones. In the sensitivity comparison, the sensitivity of the method was higher than that of the universal nested PCR targeting the 16S rRNA and tuf genes. In the specificity evaluation, it showed no cross-reactivity with phytoplasmas from the 16SrI or 16SrXXXII groups, or with other tested areca palm pathogens and endophytic bacteria. In practical application, among 22 areca palm samples collected from Wenchang, Hainan Province, 4 samples were detected as positive; additionally, the method could detect 16SrII group phytoplasmas including Parthenium hysterophorus witches'-broom phytoplasma, Cleome rutidosperma witches'-broom phytoplasma, and peanut witches'-broom phytoplasma, demonstrating good applicability. In conclusion, the nested PCR method established in this study based on the rp gene has good specificity for the 16SrII group areca palm yellow leaf phytoplasmas, and could provide technical reference for field diagnosis and disease monitoring of the 16SrII group areca palm yellow leaf phytoplasmas.
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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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