In August 2022, we found Phyllanthus urinaria with yellows and witches’-broom symptoms, respectively, in areca yellows disease garden in Wenchang city, Hainan province, which were phytoplasma infection. In order to clarify the taxonomic status of the phytoplasma strain in the yellows leaves and witches’-broom of P. urinaria. In this study, we cloned the 16S rDNA gene and ribosomal protein (rp) gene, and analyzed the gene sequence consistency, phylogenetic tree and virtual RFLP. The results showed that the 16S rDNA fragment 1246 bp and rp gene 1212 bp were cloned of P. urinaria yellows phytoplasma, the 16S rDNA fragment 1827 bp and rp gene 1240 bp were cloned of P. urinaria witches’-broom phytoplasma. The gene sequence consistency showed that the 16S rDNA gene sequence of the P. urinaria yellows phytoplasma and the P. urinaria witches’-broom phytoplasma was more than 98% consistent with the phytoplasma of 16SrⅠ group, and 100% consistent with the 16S rDNA sequence of areca yellows phytoplasma Hainan strain; the consistency of them in rp gene sequences was more than 99% to those of the rpⅠ group. The phylogenetic tree analysis showed that the 16S rDNA genes of P. urinaria yellows phytoplasma and P. urinaria witches’-broom phytoplasma were clustered in a large branch with those of aster yellows phytoplasma group (16SrⅠ), and clustered in the same small branch with those of areca yellows phytoplasma of 16SrⅠ-B subgroup, with the close genetic relationship; in the phylogenetic tree of rp gene, they were clustered in a large branch with the aster yellows phytoplasma group (rp Ⅰ), and clustered in the same small branch with those of aster yellows phytoplasma of rpⅠ-B subgroup, with the close genetic relationship. The virtual RFLP analysis showed that the RFLP map obtained by the 16S rDNA gene of the P. urinaria yellows phytoplasma and P. urinaria witches’-broom phytoplasma was the same as the reference map of the onion yellows phytoplasma of 16SrⅠ-B, and the similarity coefficient was 1.00. In summary, the P. urinaria yellows phytoplasma and the P. urinaria witches’-broom phytoplasma belong to 16SrⅠ-B subgroup in classification. The results of this study could provide a theoretical basis for the prevention and control of areca palm yellow disease by eradicating its intermediate host.
| 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 |