Molecular detection of phytoplasmas in witches’ broom diseased Ziziphus oenoplia from Andhra Pradesh, India

Ziziphus oenoplia is a wild jujube evergreen small tropical fruit tree that grows along the roadsides. During a survey in 2021, this jujube trees exhibited severe witches’ broom symptoms, therefore, an investigation was carried out, to verify the presence of phytoplasmas in the symptomatic plants. Total DNA was extracted from symptomatic leaves using a CTAB method and a PCR assay was performed using 16S rRNA gene specific primer pairs P1/P7 and R16F2n/R16R2. Amplicons of about 1.2 kb were consistently amplified from all the symptomatic plant samples in nested PCR assays, while no amplification was observed in non-symptomatic samples. Virtual RFLP analysis of 16S rDNA sequences allowed the classification of this strain into 16SrV group. The phylogenetic analysis of 16Sr DNA sequence of the detected phytoplasma strain indicates its close clustering with ‘Candidatus Phytoplasma balanitae’ and ‘Ca. P. ziziphi’ and further analyses on other housekeeping gene are necessary to define its affiliation.