First report of a 16SrII-D phytoplasma associated with phyllody in Heliopsis helianthoides

During 2017–18 surveys for phytoplasma disease presence performed in scattered green areas and parks in the Yazd province, Iran, a phyllody disease of Heliopsis helianthoides was observed. Samples from symptomatic plants were amplified using P1/P7 and R16F2n/R16R2 primers and produced the expected fragments of about 1,800 and 1,200 bp, respectively. RFLP analysis on the 1,200 bp amplicons using Kpn I, Hae III, Rsa I, Alu I, Hpa II, Hha I, Sau 3AI, and Taq I restriction enzymes showed identical profiles in all the samples tested. After their sequencing 100% identity was present among all amplicons therefore only one sequence was deposited in GenBank database (accession number MK558919). Results of the phylogenetic analysis showed the clustering of the detected H. helianthoides phyllody phytoplasma with strains related to ‘ Candidatus Phytoplasma australasia’. Real and in silico RFLP analyses allowed to classify the phytoplasma in the 16SrII-D subgroup. This is the first report of a phytoplasma associated with the H. helianthoides phyllody disease.