First Report of Desmodium styracifolium as a Novel Host for ‘Candidatus Phytoplasma australasiaticum’—Related Strains in China

<i>Desmodium styracifolium</i> (Osb.) Merr., a member of the <i>Leguminosae</i> family, is an important medicinal plant widely used in traditional Chinese medicine. In September 2024, <i>D. styracifolium</i> plants exhibiting symptoms of little leaf and stunted growth were observed in a field of Zhanjiang, Guangdong province, China. Since the symptoms resembled those associated with phytoplasma infections, total DNA was extracted from the leaves of four symptomatic plants and one healthy plant for molecular identification. Universal primer pairs (P1/P7, R16mF2/mR1) for phytoplasma detection were used to amplify the 16S rDNA fragments (~1.8 kb and ~1.4 kb), while a specific primer pair secY-F/secY-R was employed to amplify a ~1.4 kb segment of the <i>secY</i> gene. Target fragments were successfully amplified from all symptomatic samples but not from the healthy control. These amplicons were cloned and sequenced. The obtained <i>16S rDNA</i> sequence of <i>D. styracifolium</i> little leaf phytoplasma (DsLFP-GDZJ) showed the highest identity (99.67-100%) with strains of '<i>Ca.</i> Phytoplasma australasiaticum' (subgroup 16SrII-A and 16SrII-D). Phylogenetic analysis also indicated that DsLFP-GDZJ formed a small evolutionary branch with strains of '<i>Ca.</i> Phytoplasma australasiaticum' (subgroup 16SrII-A and 16SrII-D). Virtual RFLP (restriction fragment length polymorphism) analysis of the 16S <i>rDNA</i> sequence demonstrated DsLFP-GDZJ belongs to the 16SrII-A subgroup (GenBank accession number L33765). The <i>secY</i> gene sequence of DsLFP-GDZJ also showed the highest similarity and the closest relationship with those of the 16SrII-A subgroup phytoplasma strains. These results showed that DsLFP-GDZJ is a strain of '<i>Ca.</i> Phytoplasma australasiaticum' (16SrII-A subgroup). To our knowledge, this is the first report of '<i>Ca.</i> Phytoplasma australasiaticum'-related phytoplasma associated with <i>D. styracifolium</i> little leaf disease in China, thereby establishing <i>D. styracifolium</i> (Osb.) Merr. as a new host plant of phytoplasma.