During the year 2017, surveys were carried out in strawberry fields in Biobío and Araucanía regions in Chile, where strawberry plants with symptoms of phyllody, achenes’ hypertrophy and leaf reddening had been observed. In the symptomatic plants, two phytoplasmas belonging to the ribosomal subgroups 16SrXIII-F and 16SrXIII-K have been identified. In order to obtain specific molecular tools for the detection of these phytoplasmas, two regions containing the genes encoding the ribosomal proteins 12p and 36p were used, and the phylogenetic trees obtained unambiguously support the distinctness of the two ribosomal subgroups.