During field surveys conducted in 2024 and 2025 in pepper production areas in Altunhisar distinct of Niğde province, Türkiye, plants showing phytoplasma-like symptoms were observed. A total of 25 symptomatic pepper samples were collected, and total nucleic acids were extracted using the CTAB method. The samples were analyzed for phytoplasma infection by PCR amplification of the 16S rRNA gene region. In addition, two asymptomatic pepper plants were included as negative controls. PCR analysis revealed that all samples showing symptoms produced amplicons of the expected size, while no amplification was obtained from the asymptomatic plants. Sequencing and phylogenetic analyses revealed that the detected phytoplasma belonged to the 16SrVI-A subgroup, corresponding to ‘Candidatus Phytoplasma trifolii’. The subgroup assignment was further confirmed by in silico (virtual) RFLP analysis of the 16S rRNA gene. The representative 16S rRNA gene sequence of one strain obtained in this study was deposited in the GenBank database (GenBank accession number PX885052). Based on the available literature, this study provides the first molecular evidence of ‘Candidatus Phytoplasma trifolii’ infection in pepper plants grown in Niğde province, Türkiye. The findings contribute to the current knowledge on the distribution of pepper-infecting phytoplasmas in Türkiye and emphasize the need for further epidemiological and vector-related studies.