Phytoplasma disease of tamarisk witches' broom in the Chah Afzal region of Yazd province has a great importance due to the widespread and damage to tamarisk trees. The tamarisk trees are planted in Chah Afzal as windbreaks for pistachio orchards, and the extensive spread of this phytoplasma disease turns them into a source of contamination for other hosts including pistachios. The infected tamarisk trees showed phytoplasma disease symptoms, including yellowing, leaf reduction, and witches' broom were observed and investigated during 2019 to 2022. Plant samples with suspicious symptoms of phytoplasma were collected, and after extracting the total DNA of the samples, a one-step polymerase chain reaction (PCR) (using the primer pair P1/P7) or two-step PCR (using the primer pairs P1/P7 and then R16mF2/R16mR2 and R16F2n/R16R2) were performed. The results of this research indicated the presence of phytoplasma subgroup D associated with tamarisk witches' broom disease. Phytoplasma of the peanut witches' broom group have been previously reported in pistachios, suggesting that pistachios also can be host of this phytoplasma group. Salt-tolerant plants like tamarisk are grown as windbreaks and soil erosion reducers in most farms, orchards, and adjacent areas in the arid and semi-arid regions of Yazd province characterized by high evaporation and transpiration rates and low rainfall. Tamarisk plants can serve as a potential source of reservoir for phytoplasma and their vectors, contributing to the contamination of pistachio trees and other agricultural and natural resources plants with peanut witches' broom phytoplasma.