Title: Association of 16SrII-C phytoplasma with lentil phyllody disease in Pakistan
Abstract: In April 2011, lentil (Lens culinaris) plants were found with symptoms reminiscent of phytoplasma infection for the first time in Pakistan. Floral malformation, chlorosis, little leaf and extensive proliferation of branches were the most common symptoms. For the confirmation of phytoplasma, total DNA was extracted separately from eight symptomatic plants and was amplified by nested PCR using universal 16Sr DNA phytoplasma primers P1/P7. One representative PCR product was sequenced in both directions using primers R16F2n/R16R2 and was shown to have 100 % identity to phytoplasmas of the 16SrII-C subgroup. The infectious agent was successfully transmitted both by grafting and by the leafhopper Orosius albicinctus, while transmission via seed failed to produce disease symptoms under glasshouse conditions. Infection of lentils with phytoplasmas has not previously been reported in the literature, so to our knowledge, this is the first report of phytoplasma from the 16SrII-C group naturally infecting lentil in Pakistan.